2-propynyl adenosine analogs having A2A agonist activity and compositions thereof

ABSTRACT

The invention provides compounds having the following general formula (I):  
                 
 
     wherein X, R 1 , R 2 , R 7  and Z are as described here.

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority of U.S. provisional patentapplication Serial No. 60/326,517, filed Oct. 1, 2001, and U.S.provisional patent application Serial No. 60/383,200, filed May 24,2001, both of which are incorporated by reference herein.

GOVERNMENT FUNDING

[0002] The invention described herein was made with government supportunder Grant Number (RO1-HL37942), awarded by the National ScienceFoundation. The United States Government has certain rights in theinvention.

BACKGROUND OF THE INVENTION

[0003] The inflammatory response serves the purpose of eliminatingharmful agents from the body. There is a wide range of pathogenicinsults that can initiate an inflammatory response including infection,allergens, autoimmune stimuli, immune response to transplanted tissue,noxious chemicals, and toxins, ischemia/reperfusion, hypoxia, mechanicaland thermal trauma. Inflammation normally is a very localized actionwhich serves in expulsion, attenuation by dilution, and isolation of thedamaging agent and injured tissue. The body's response becomes an agentof disease when it results in inappropriate injury to host tissues inthe process of eliminating the targeted agent, or responding to atraumatic insult.

[0004] As examples, inflammation is a component of pathogenesis inseveral vascular diseases or injuries. Examples include:ischemia/reperfusion injury (N. G. Frangogiannis et al., in MyocardialIschemia: Mechanisms, Reperfusion, Protection, M. Karmazyn, ed.,Birkhuser Verlag (1996) at 236-284; H. S. Sharma et al., Med. ofInflamm., 6, 175 (1987)), atherosclerosis (R. Ross, Nature, 362, 801(1993)), inflammatory aortic aneurysms (N. Girardi et al., Ann. Thor.Surg., 64, 251 (1997); D. I. Walker et al., Brit. J. Surg., 59, 609(1972); R. L. Pennell et al., J. Vase. Surg., 2, 859 (1985)), andrestenosis following balloon angioplasty (see, R. Ross cited above). Thecells involved with inflammation include leukocytes (i.e., the immunesystem cells—neutrophils, cosinophils, lymphocytes, monocytes,basophils, macrophages, dendritic cells, and mast cells), the vascularendothelium, vascular smooth muscle cells, fibroblasts, and myocytes.

[0005] The release of inflammatory cytokines such as tumor necrosisfactor-alpha (TNFα) by leukocytes is a means by which the immune systemcombats pathogenic invasions, including infections. TNFα stimulates theexpression and activation of adherence factors on leukocytes andendothelial cells, primes neutrophils for an enhanced inflammatoryresponse to secondary stimuli and enhances adherent neutrophil oxidativeactivity. See, Sharma et al., cited above. In addition,macrophages/dendritic cells act as accessory cells processing antigenfor presentation to lymphocytes. The lymphocytes, in turn, becomestimulated to act as pro-inflammatory cytotoxic cells.

[0006] Generally, cytokines stimulate neutrophils to enhance oxidative(e.g., superoxide and secondary products) and nonoxidative (e.g.,myeloperoxidase and other enzymes) inflammatory activity. Inappropriateand over-release of cytokines can produce counterproductive exaggeratedpathogenic effects through the release of tissue-damaging oxidative andnonoxidative products (K. G. Tracey et al., J. Exp. Med., 167, 1211(1988); and D. N. Männel et al., Rev. Infect. Dis., 9 (suppl. 5),S602-S606 (1987)). For example, TNFα can induce neutrophils to adhere tothe blood vessel wall and then to migrate through the vessel to the siteof injury and release their oxidative and non-oxidative inflammatoryproducts.

[0007] Although monocytes collect slowly at inflammatory foci, givenfavorable conditions, the monocytes develop into long-term residentaccessory cells and macrophages. Upon stimulation with an inflammationtrigger, monocytes/macrophages also produce and secrete an array ofcytokines (including TNFα), complement, lipids, reactive oxygen species,proteases and growth factors that remodel tissue and regulatesurrounding tissue functions.

[0008] For example, inflammatory cytokines have been shown to bepathogenic in: arthritis (C. A. Dinarello, Semin. Immunol., 4 133(1992)); ischemia (A. Seekamp et al., Agents-Actions-Supp., 41, 137(1993)); septic shock (D. N. Männel et al., Rev. Infect. Dis., 9 (suppl.5), S602-S606 (1987)); asthma (N. M. Cembrzynska et al., Am. Rev.Respir. Dis., 147, 291 (1993)); organ transplant rejection (D. K.Imagawa et al., Transplantation, 51, 57 (1991); multiple sclerosis (H.P. Hartung, Ann. Neurol., 33, 591 (1993)); AIDS (T. Matsuyama et al.,AIDS, 5, 1405 (1991)); and in alkali-burned eyes (F. Miyamoto et al.,Opthalmic Res., 30, 168 (1997)). In addition, superoxide formation inleukocytes has been implicated in promoting replication of the humanimmunodeficiency virus (HIV) (S. Legrand-Poels et al., AIDS Res. Hum.Retroviruses, 6, 1389 (1990)).

[0009] It is well known that adenosine and some analogs of adenosinethat nonselectively activate adenosine receptor subtypes decreaseneutrophil production of inflammatory oxidative products (B. N.Cronstein et al., Ann. N.Y. Acad. Sci., 451, 291 (1985); P. A. Robertset al., Biochem. J., 227, 669 (1985); D. J. Schrier et al., J. Immunol.,137, 3284 (1986); B. N. Cronstein et al., Clinical Immunol. andImmunopath., 42, 76 (1987); M. A. Iannone et al., in Topics andPerspective in Adenosine Research, E. Gerlach et al., eds.,Springer-Verlag, Berlin, p. 286 (1987); S. T. McGarrity et al., J.Leukocyte Biol., 44, 411421 (1988); J. De La Harpe et al., J. Immunol.,143, 596 (1989); S. T. McGarrity et al., J. Immunol., 142, 1986 (1989);and C. P. Nielson et al., Br. J. Pharmacol., 97, 882 (1989)). Forexample, adenosine has been shown to inhibit superoxide release fromneutrophils stimulated by chemoattractants such as the synthetic mimicof bacterial peptides, f-met-leu-phe (fMLP), and the complementcomponent C₅a (B. N. Cronstein et al., J. Immunol., 135, 1366 (1985)).Adenosine can decrease the greatly enhanced oxidative burst of PMN(neutrophil) first primed with TNF-α and then stimulated by a secondstimulus such as f-met-leu-phe (G. W. Sullivan et al., Clin. Res., 41,172A (1993)). Additionally, it has been reported that adenosine candecrease the rate of HIV replication in a T-cell line (S. Sipka et al.,Acta. Biochim. Biopys. Hung., 23, 75 (1988)). However, there is noevidence that in vivo adenosine has anti-inflammatory activity (G. S.Firestein et al., Clin. Res., 41, 170A (1993); and B. N. Cronstein etal., Clin. Res., 41, 244A (1993)).

[0010] It has been suggested that there is more than one subtype ofadenosine receptor on neutrophils that can have opposite effects onsuperoxide release (B. N. Cronstein et al., J. Clin. Invest., 85, 1150(1990)). The existence of A_(A2) receptor on neutrophils was originallydemonstrated by Van Calker et al. (D. Van Calker et al., Eur. J.Pharmacology, 206, 285 (1991)).

[0011] There has been progressive development of compounds that are moreand more potent and/or selective as agonists of A_(A2) adenosinereceptors (AR) based on radioligand binding assays and physiologicalresponses. Initially, compounds with little or no selectivity for A_(A2)receptors were developed, such as adenosine itself or 5′-carboxamides ofadenosine, such as 5′-N-ethylcarboxamidoadenosine (NECA) (B. N.Cronstein et al., J. Immunol., 135, 1366 (1985)). Later, it was shownthat addition of 2-alkylamino substituents increased potency andselectivity, e.g., CV1808 and CGS21680 (M. F. Jarvis et al., J.Pharmacol. Exp. Ther., 251, 888 (1989)). 2-Alkoxy-substituted adenosinederivatives such as WRC-0090 are even more potent and selective asagonists at the coronary artery A_(A2) receptor (M. Ueeda et al., J.Med. Chem., 34, 1334 (1991)). The 2-alklylhydrazino adenosinederivatives, e.g., SHA 211 (also called WRC-0474) have also beenevaluated as agonists at the coronary artery A_(A2) receptor (K. Niiyaet al., J. Med. Chem., 35, 4557 (1992)).

[0012] There is one report of the combination of relatively nonspecificadenosine analogs, R-phenylisopropyladenosine (R-PIA) and2-chloroadenosine (Cl-Ado) with a phosphodiesterase (PDE) inhibitorresulting in a lowering of neutrophil oxidative activity (M. A. Iannoneet al., Topics and Perspectives in Adenosine Research, E. Garlach etal., eds., Springer-Verlag, Berlin, pp. 286-298 (1987)). However, R-PIAand Cl-Ado analogs are actually more potent activators of A₁ adenosinereceptors than of A_(A2) adenosine receptors and, thus, are likely tocause side effects due to activation of A₁ receptors on cardiac muscleand other tissues causing effects such as “heart block.”

[0013] R. A. Olsson et al. (U.S. Pat. No. 5,278,150) disclose selectiveadenosine A₂ receptor agonists of the formula:

[0014] wherein Rib is ribosyl, R₁ can be H and R₂ can be cycloalkyl. Thecompounds are disclosed to be useful for treating hypertension,atherosclerosis and as vasodilators.

[0015] Olsson et al. (U.S. Pat. No. 5,140,015) disclose certainadenosine A₂ receptor agonists of formula:

[0016] wherein C(X)BR₂ can be CH₂OH and R₁ can be alkyl- or alkoxyalkyl.The compounds are disclosed to be useful as vasodilators or anantihypertensives.

[0017] Linden et al. (U.S. Pat. No. 5,877,180) is based on the discoverythat certain inflammatory diseases, such as arthritis and asthma, may beeffectively treated by the administration of compounds which areselective agonists of A_(2A) adenosine receptors, preferably incombination with a Type IV phosphodiesterase inhibitor. An embodiment ofthe Linden et al. invention provides a method for treating inflammatorydiseases by administering an effective amount of an A_(2A) adenosinereceptor of the following formula:

[0018] wherein R and X are as described in the patent.

[0019] In one embodiment, the Linden et al. invention involves theadministration of a Type IV phosphodiesterase (PDE) inhibitor incombination with the A_(2A) adenosine receptor agonist. The Type IVphosphodiesterase (PDE) inhibitor includes racemic and optically active4-(polyalkoxyphenyl)-2-pyrrolidones of the following formula:

[0020] wherein R′, R¹⁸, R¹⁹ and X are as disclosed and described in U.S.Pat. No. 4,193,926. Rolipram is an example of a suitable Type IV PDEinhibitor included within the above formula.

[0021] G. Cristalli (U.S. Pat. No. 5,593,975) discloses 2-arylethynyl,2-cycloalkylethynyl or 2-hydroxyalkylethynyl derivatives, wherein theriboside residue is substituted by carboxy amino, or substituted carboxyamino (R₃HNC(O)—). 2-Alkynylpurine derivatives have been disclosed inMiyasaka et al. (U.S. Pat. No. 4,956,345), wherein the 2-alkynyl groupis substituted with (C₃-C₁₆)alkyl. The '975 compounds are disclosed tobe vasodilators and to inhibit platelet aggregation, and thus to beuseful as anti-ischemic, anti-atherosclerosis and anti-hypertensiveagents.

[0022] Recently, U.S. Pat. No. 6,232,297 to Linden, et al. disclosedcompounds having the general formula:

[0023] wherein each R is H, X is ethylaminocarbonyl and R¹ is4-carboxycyclo-hexylmethyl (DWH-146a), R¹ is4-methoxycarbonylcyclohexylmethyl (DWH-146e) or R¹ is4-acetoxymethyl-cyclohexylmethyl (JMR-193). These compounds are reportedto be A_(A2) agonists.

[0024] However, a continuing need exists for selective A₂ adenosinereceptor agonists useful for therapeutic applications, that have reducedside effects.

SUMMARY OF THE INVENTION

[0025] The present invention comprises compounds and methods of theiruse for the treatment of inflammatory activity in mammalian tissue. Theinflammatory tissue activity can be due to pathological agents or can bedue to physical, chemical or thermal trauma, or the trauma of medicalprocedures, such as organ, tissue or cell transplantation, angioplasty(PCTA), inflammation following ischemia/reperfusion, or grafting. Thepresent compounds comprise a novel class of 2-alkynyladenosinederivatives, substituted at the ethyn-2-yl position by substitutedcycloalkyl and heterocycle (heterocyclic) moieties. Preferably, theriboside residue is substituted at the 5′-position by an N-alkyl-(orcycloalkyl)carboxyamino (“aminocarbonyl”) moiety (“X”). Thus, thepresent invention provides a method for inhibiting the inflammatoryresponse in a mammal, such as a human subject, and protecting the tissuesubject to the response, by administering an effective amount of one ormore compounds of the invention.

[0026] The compounds of the invention have general formula (I):

[0027] wherein

[0028] Z is CR³R⁴R⁵ or NR⁴R⁵;

[0029] each R₁ is independently hydrogen, halo, —OR^(a), —SR^(a),(C₁-C₈)alkyl, cyano, nitro, trifluoromethyl, trifluoromethoxy,C₃₋₈cycloalkyl, heterocycle, hetrocycle(C₁-C₈)alkylene-, aryl,aryl(C₁-C₈)alkylene-, heteroaryl, heteroaryl(C₁-C₈)alkylene-, -CO₂R^(a),R^(a)C(═O)O—, R^(a)C(═O)—, —OCO₂R^(a), R^(a)R^(b)NC(═O)O—,R^(b)OC(═O)N(R^(a))—, R^(a)R^(b)N—, R^(a)R^(b)NC(═O)—,R^(a)C(═O)N(R^(b))—, R^(a)R^(b)NC(═O)N(R^(b))—,R^(a)R^(b)NC(═S)N(R^(b))—, —OPO₃R^(a), R^(a)OC(═S)—, R^(a)C(═S)—,—SSR^(a), R^(a)S(═O)—, R^(a)S(═O)₂—, —N═NR^(a), or —OPO₂R^(a);

[0030] each R² is independently hydrogen, halo, (C₁-C₈)alkyl,(C₃-C₈)cycloalkyl, heterocycle, heterocycle(C₁-C₈)alkylene-, aryl,aryl(C₁-C₈)alkylene-, heteroaryl, or heteroaryl(C₁-C₈)alkylene-; or

[0031] R¹ and R² and the atom to which they are attached is C═O, C═S orC═NR^(c).

[0032] R⁴ and R⁵ together with the atoms to which they are attached forma saturated or partially unsaturated, mono-, bicyclic- or aromatic ringhaving 3, 4, 5, 6, 7, 8, 9 or 10 ring atoms optionally comprising 1, 2,3, or 4 heteroatoms selected from non-peroxide oxy (—O—), thio (—S—),sulfinyl (—SO—), sulfonyl (—S(O)₂—) or amine (—NR^(a)—) in the ring;

[0033] wherein any ring comprising R⁴ and R⁵ is substituted with from 1to 14 R⁶ groups; wherein each R⁶ is independently halo, —OR^(a),—SR^(a), (C₁-C₈)alkyl, cyano, nitro, trifluoromethyl, trifluoromethoxy,(C₁-C₈)cycloalkyl, (C₆-C₁₂)bicycloalkyl, heterocycle or hetrocycle(C₁-C₈)alkylene-, aryl, aryl (C₁-C₈)alkylene-, heteroaryl,heteroaryl(C₁-C₈)alkylene-, —CO₂R^(a), R^(a)C(═O)O—, R^(a)C(═O)—,—OCO₂R^(a), R^(a)R^(b)NC(═O)O—, R^(b)OC(═O)N(R^(a))—, R^(a)R^(b)N—,R^(a)R^(b)NC(═O)—, R^(a)C(═O)N(R^(b))—, R^(a)R^(b)NC(═O)N(R^(b))—,R^(a)R^(b)NC(═S)N(R^(b))—, —OPO₃R^(a), R^(a)OC(═S)—, R^(a)C(═S)—,—SSR^(a), R^(a)S(═O)—, —NNR^(a),—OPO₂R^(a), or two R⁶ groups and theatom to which they are attached is C═O, C═S or; two R⁶ groups togetherwith the atom or atoms to which they are attached can form a carbocyclicor heterocyclic ring.

[0034] R³ is hydrogen, halo, —OR^(a), —SR^(a), (C₁-C₈)alky1, cyano,nitro, trifluoromethyl, trifluoromethoxy, (C₃-C₈)cycloalkyl,heterocycle, hetrocycle(C₁-C₈)alkylene-, aryl, aryl(C₁-C₈)alkylene-,heteroaryl, heteroaryl(C₁-C₈)alkylene-, —CO₂R^(a), R^(a)C(═O)O—,R^(a)C(═O)—, —OCO₂R^(a), R^(a)R^(b)NC(═O)O—, R^(b)OC(═O)N(R^(a))—,R^(a)R^(b)N—, R^(a)R^(b)NC(═O)—, R^(a)C(═O)N(R^(b))—,R^(a)R^(b)NC(═O)N(R^(b))—, R^(a)R^(b)NC(═S)N(R^(b))—, —OPO₃R^(a),R^(a)OC(═S)—, R^(a)C(═S)—, —SSR^(a), R^(a)S(═O)—, R^(a)S(═O)₂—,—NNR^(a), —OPO₂R^(a); or if the ring formed from CR⁴R⁵ is aryl orhetreroaryl or partially unsaturated then R³ can be absent;

[0035] each R⁷is independently hydrogen, (C₁-C₈)alkyl,(C₃-C₈)cycloalkyl, aryl or aryl(C₁-C₈)alkylene, heteroaryl,heteroaryl(C₁-C₈)alkylene-;

[0036] X is —CH₂OR^(a), —CO₂R^(a), —OC(O)R^(a), —CH₂OC(O)R^(a),—C(O)NR^(a)R^(b), —CH₂SR^(a), —C(S)OR^(a), —OC(S)R^(a), —CH₂OC(S)R^(a)or C(S)NR^(a)R^(b) or —CH₂N(R^(a))(R^(b));

[0037] wherein any of the alkyl, cycloalkyl, heterocycle, aryl, orheteroaryl, groups of R¹, R², R³, R⁶ and R⁷is optionally substituted oncarbon with one or more (e.g. 1, 2, 3, or 4) substituents selected fromthe group consisting of halo, —OR^(a), —SR^(a), (C₁-C₈)alkyl, cyano,nitro, trifluoromethyl, trifluoromethoxy, (C₃-C₈)cycloalkyl,(C₆-C₁₂)bicycloalkyl, heterocycle or hetrocycle(C₁-C₈)alkylene-, aryl,aryloxy, aryl (C₁-C₈)alkylene-, heteroaryl, heteroaryl(C₁-C₈)alkylene-,—CO₂R^(a), R^(a)C(═O)O—, R^(a)C(═O)—, —OCO₂R^(a), R^(a)R^(b)NC(═O)O—,R^(b)OC(═O)N(R^(a))—, R^(a)R^(b)N—, R^(a)R^(b)NC(═O)—,R^(a)C(═O)N(R^(b))—, R^(a)R^(b)NC(═O)N(R^(b))—,R^(a)R^(b)NC(═S)N(R^(b))—, —OPO₃R^(a), R^(a)OC(═S)—, R^(a)C(═S)—,—SSR^(a), R^(a)S(═O)_(p)—, R^(a)R^(b)NS(O)_(p)—, N═NR^(a), and—OPO₂R^(a);

[0038] wherein any (C₁-C₈)alkyl, (C₃-C₈)cycloalkyl,(C₆-C₁₂)bicycloalkyl, (C₁-C₈)alkoxy, (C₁-C₈)alkanoyl, (C₁-C₈)alkylene,or heterocycle, is optionally partially unsaturated;

[0039] R^(a) and R^(b) are each independently hydrogen, (C₁-C₈)alkyl, or(C₁-C₈)alkyl substituted with 1-3 (C₁-C₈)alkoxy, (C₃-C₈)cycloalkyl,(C₁-C₈)alkylthio, amino acid, aryl, aryl(C₁-C₈)alkylene, heteroaryl, orheteroaryl(C₁-C₈)alkylene; or R^(a) and R^(b), together with thenitrogen to which they are attached, form a pyrrolidino, piperidino,morpholino, or thiomorpholino ring; and

[0040] R^(c) is hydrogen or (C₁-C₆)alkyl;

[0041] m is 0 to about 8 and p is 0 to 2;

[0042] provided that when CR⁴R⁵ is a carbocyclic ring then at least oneof R¹, R², or R³ is a group other than hydrogen or at least one R⁶ groupis a group other than —CH₂OH, —CO₂R^(a), R^(a)C(═O)O—, R^(a)C(═O)OCH₂—or R^(a)R^(b)NC(═O)—;

[0043] provided that m is at least 1 when Z is NR⁴R⁵;

[0044] or a pharmaceutically acceptable salt thereof.

[0045] The invention provides a compound of formula I for use in medicaltherapy, preferably for use in treating inflammation or protectingmammalian tissue from inflammation such as an inflammatory response,e.g., resulting from allergy, trauma or ischemia/reperfusion injury, aswell as the use of a compound of formula I for the manufacture of amedicament for the treatment of an inflammatory response due to apathological condition or symptom in a mammal, such as a human, which isassociated with inflammation.

[0046] Although certain A_(A2) adenosine receptor agonists have beenreported to be vasodilators, and thus to be useful to directly treathypertension, thrombus, atherosclerosis and the like, thetissue-protective anti-inflammatory activity of the compounds of formula(I) is not suggested by the prior art.

[0047] The invention also includes the use of a combination of thesecompounds with type IV phosphodiesterase inhibitors to preferably causesynergistic decreases in the inflammatory response mediated byleukocytes.

[0048] The invention also provides a pharmaceutical compositioncomprising an effective amount of the compound of formula I, or apharmaceutically acceptable salt thereof, in combination with apharmaceutically acceptable diluent or carrier, and optionally, incombination with a Type IV phosphodiesterase (PDE) inhibitor.Preferably, the composition is presented as a unit dosage form.

[0049] Additionally, the invention provides a therapeutic method forpreventing or treating a pathological condition or symptom in a mammal,such as a human, wherein the activity of A_(A2) adenosine receptors isimplicated and agonism of said receptors is desired, comprisingadministering to a mammal in need of such therapy, an effective amountof a compound of formula I, or a pharmaceutically acceptable saltthereof. It is believed that activation of A_(A2) adenosine receptorsinhibits inflammation by affecting neutrophils, mast cells,monocytes/macrophages, platelets T-cells and/or eosinophils. Inhibitionof these inflammatory cells results in tissue protection followingtissue insults.

[0050] Among the inflammatory responses that can be treated (includingtreated prophylactically) with a compound of formula I, optionally witha Type IV PDE inhibitor, are inflammation due to:

[0051] (a) autoimmune stimulation (autoimmune diseases), such as lupuserythematosus, multiple sclerosis, infertility from endometriosis, typeI diabetes mellitus including the destruction of pancreatic isletsleading to diabetes and the inflammatory consequences of diabetes,including leg ulcers, Crohn's disease, ulcerative colitis, inflammatorybowel disease, osteoporosis and rheumatoid arthritis;

[0052] (b) allergic diseases such as asthma, hay fever, rhinitis, poisonivy, vernal conjunctivitis and other eosinophil-mediated conditions;

[0053] (c) skin diseases such as psoriasis, contact dermatitis, eczema,infectious skin ulcers, open wounds, cellulitis;

[0054] (d) infectious diseases including sepsis, septic shock,encephalitis, infectious arthritis, endotoxic shock, gram negativeshock, Jarisch-Herxheimer reaction, anthrax, plague, tularemia, ebola,shingles, toxic shock, cerebral malaria, bacterial meningitis, acuterespiratory distress syndrome (ARDS), lyme disease, HIV infection,(TNFα-enhanced HIV replication, TNFα inhibition of reverse transcriptaseinhibitor activity);

[0055] (e) wasting diseases: cachexia secondary to cancer and HIV;

[0056] (f) organ, tissue or cell transplantation (e.g., bone marrow,cornea, kidney, lung, liver, heart, skin, pancreatic islets) includingtransplant rejection, and graft versus host disease;

[0057] (g) adverse effects from drug therapy, including adverse effectsfrom amphotericin B treatment, adverse effects from immunosuppressivetherapy, e.g., interleukin-2 treatment, adverse effects from OKT3treatment, contrast dyes, antibiotics, adverse effects from GM-CSFtreatment, adverse effects of cyclosporine treatment, and adverseeffects of aminoglycoside treatment, stomatitis and mucositis due toimmunosuppression;

[0058] (h) cardiovascular conditions including circulatory diseasesinduced or exasperated by an inflammatory response, such as ischemia,atherosclerosis, peripheral vascular disease, restenosis followingangioplasty, inflammatory aortic aneurysm, vasculitis, stroke, spinalcord injury, congestive heart failure, hemorrhagic shock,ischemia/reperfusion injury, vasospasm following subarachnoidhemorrhage, vasospasm following cerebrovascular accident, pleuritis,pericarditis, and the cardiovascular complications of diabetes;

[0059] (i) dialysis, including pericarditis, due to peritoneal dialysis;

[0060] (j) gout; and

[0061] (k) chemical or thermal trauma due to burns, acid, alkali and thelike.

[0062] Of particular interest and efficacy is the use of the presentcompounds to limit inflammatory responses where the ischemia/reperfusioninjury is caused by angioplasty or throbolysis. Also of particularinterest and efficacy is the use of the present compounds to limitinflammatory responses due to organ, tissue or cell transplantation,i.e., the transplantation of allogeneic or xenogeneic tissue into amammalian recipient, autoimmune diseases and inflammatory conditions dueto circulatory pathologies and the treatment thereof, includingangioplasty, stent placement, shunt placement or grafting. Unexpectedly,it was found that administration of one or more compounds of formula (I)was effective after the onset of the inflammatory response, e.g., afterthe subject was afflicted with the pathology or trauma that initiatesthe inflammatory response.

[0063] Tissue or cells comprising ligand bound receptor sites can beused to measure the selectively of test compounds for specific receptorsubtypes, the amount of bioactive compound in blood or otherphysiological fluids, or can be used as a tool to identify potentialtherapeutic agents for the treatment of diseases or conditionsassociated with receptor site activation, by contacting said agents withsaid ligand-receptor complexes, and measuring the extent of displacementof the ligand and/or binding of the agent, or the cellular response tosaid agent (e.g., cAMP accumulation).

BRIEF DESCRIPTION OF THE FIGURES

[0064]FIG. 1 illustrates the results of a comparison of the depressionof blood pressure in rats using the compound ATL-146e and JR4007 at 100ug/kg.

[0065]FIG. 2 illustrates the results of a dose-response experiment forthe depression of blood pressure in rats using the compound JR4007 atconcentrations of 1, 10, and 100 ug/kg.

[0066]FIG. 3 illustrates the results of a comparison of the depressionof blood pressure in rats using test compounds at 1 ug/kg.

[0067]FIG. 4 illustrates the results of a comparison of the depressionof blood pressure in rats using test compound JR 3223 in two animals.

[0068]FIG. 5 illustrates the results of a of a dose-response experimentfor the depression of blood pressure in rats using for JR4051 atconcentrations of 1 and 10 ug/kg.

[0069]FIG. 6 illustrates the results of a comparison of the depressionof blood pressure in rats using the compounds of the invention.

[0070] FIGS. 7-16 illustrate the results of the coronary blood flow fortest compounds in dogs.

[0071]FIG. 17 illustrates the results of the liver ischemia/reperfusioninjury test

DETAILED DESCRIPTION OF THE INVENTION

[0072] The following definitions are used, unless otherwise described.Halo is fluoro, chloro, bromo, or iodo. Alkyl, alkoxy, aralkyl,alkylaryl, etc. denote both straight and branched alkyl groups; butreference to an individual radical such as “propyl” embraces only thestraight chain radical, a branched chain isomer such as “isopropyl”being specifically referred to. Aryl includes a phenyl radical or anortho-fused bicyclic carbocyclic radical having about nine to ten ringatoms in which at least one ring is aromatic. Heteroaryl encompasses aradical attached via a ring carbon of a monocyclic aromatic ringcontaining five or six ring atoms consisting of carbon and one to fourheteroatoms each selected from the group consisting of non-peroxideoxygen, sulfur, and N(X) wherein X is absent or is H, O, (C₁-C₄)alkyl,phenyl or benzyl, as well as a radical of an ortho-fused bicyclicheterocycle of about eight to ten ring atoms derived therefrom,particularly a benz-derivative or one derived by fusing a propylene,trimethylene, or tetramethylene diradical thereto.

[0073] It will be appreciated by those skilled in the art that thecompounds of formula (I) have more than one chiral center and may beisolated in optically active and racemic forms. Preferably, the ribosidemoiety of formula (I) is derived from D-ribose, i.e., the 3′,4′-hydroxylgroups are alpha to the sugar ring and the 2′ and 5′ groups is beta (3R,4S, 2R, 5S). When the two groups on the cyclohexyl group are in the 1-and 4-position, they are preferably trans. Some compounds may exhibitpolymorphism. It is to be understood that the present inventionencompasses any racemic, optically-active, polymorphic, orstereoisomeric form, or mixtures thereof, of a compound of theinvention, which possess the useful properties described herein, itbeing well known in the art how to prepare optically active forms (forexample, by resolution of the racemic form by recrystallizationtechniques, or enzymatic techniques, by synthesis from optically-activestarting materials, by chiral synthesis, or by chromatographicseparation using a chiral stationary phase) and how to determineadenosine agonist activity using the tests described herein, or usingother similar tests which are well known in the art.

[0074] Specific and preferred values listed below for radicals,substituents, and ranges, are for illustration only; they do not excludeother defined values or other values within defined ranges for theradicals and substituents.

[0075] Specifically, (C₁-C₈)alkyl can be methyl, ethyl, propyl,isopropyl, butyl, iso-butyl, sec-butyl, pentyl, 3-pentyl, hexyl, heptylor octyl. As used herein, the term “cycloalkyl” encompasses bicycloalkyl(norbornyl, 2.2.2-bicyclooctyl, etc.) and tricycloalkyl (adamantyl,etc.), optionally comprising 1-2 N, O or S. Cycloalkyl also encompasses(cycloalkyl)alkyl. Thus, (C₃-C₆)cycloalkyl can be cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl and the like. (C₁-C₈)alkoxy can bemethoxy, ethoxy, propoxy, isopropoxy, butoxy, iso-butoxy, sec-butoxy,pentoxy, 3-pentoxy, or hexyloxy; (C₂-C₆)alkenyl can be vinyl, allyl,1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 1-pentenyl,2-pentenyl, 3-pentenyl, 4-pentenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl,4-hexenyl, or 5-hexenyl; (C₂-C₆)alkynyl can be ethynyl, 1-propynyl,2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl,3-pentynyl, 4-pentynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl, or5-hexynyl; (C₁-C₆)alkanoyl can be acetyl, propanoyl or butanoyl;halo(C₁-C₆)alkyl can be iodomethyl, bromomethyl, chloromethyl,fluoromethyl, trifluoromethyl, 2-chloroethyl, 2-fluoroethyl,2,2,2-trifluoroethyl, or pentafluoroethyl; hydroxy(C₁-C₆)alkyl can behydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl, 1-hydroxypropyl,2-hydroxypropyl, 3-hydroxypropyl, 1-hydroxybutyl, 4-hydroxybutyl,1-hydroxypentyl, 5-hydroxypentyl, 1-hydroxyhexyl, or 6-hydroxyhexyl;(C₁-C₆)alkoxycarbonyl (CO₂R²) can be methoxycarbonyl, ethoxycarbonyl,propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, pentoxycarbonyl, orhexyloxycarbonyl; (C₁-C₆)alkylthio can be methylthio, ethylthio,propylthio, isopropylthio, butylthio, isobutylthio, pentylthio, orhexylthio, (C₂-C₆)alkanoyloxy can be acetoxy, propanoyloxy, butanoyloxy,isobutanoyloxy, pentanoyloxy, or hexanoyloxy; aryl can be phenyl,indenyl, or naphthyl; and heteroaryl can be furyl, imidazolyl,triazolyl, triazinyl, oxazoyl, isoxazoyl, thiazolyl, isothiazoyl,pyraxolyl, pyrrolyl, pyrazinyl, tetrazolyl, puridyl (or its N-oxide),thientyl, pyrimidinyl (or its N-oxide), indolyl, isoquinolyl (or itsN-oxide) or quinolyl (or its N-oxide).

[0076] Aryl denotes a phenyl radical or an ortho-fused bicycliccarbocyclic radical having about nine to ten ring atoms in which atleast one ring is aromatic. Heteroaryl denotes a radical of a monocyclicaromatic ring containing five or six ring atoms consisting of carbon and1, 2, 3, or 4 heteroatoms each selected from the group consisting ofnon-peroxide oxygen, sulfur, and N(Y) wherein Y is absent or is H, O,(C₁-C₈)alkyl, phenyl or benzyl, as well as a radical of an ortho-fusedbicyclic heterocycle of about eight to ten ring atoms derived therefrom,particularly a benz-derivative or one derived by fusing a propylene,trimethylene, or tetramethylene diradical thereto.

[0077] The term “heterocycle” generally represents a non aromaticheterocyclic group, having from 3 to about 10 ring atoms, which can besaturated or partially unsaturated, containing at least one heteroatom(e.g., 1, 2, or 3) selected from the group consisting of oxygen,nitrogen, and sulfur. Specific, “heterocycle” groups include monocyclic,bicyclic, or tricyclic groups containing one or more heteroatomsselected from the group consisting of oxygen, nitrogen, and sulfur. A“heterocycle” group also can include one or more oxo groups (═O)attached to a ring atom. Nonlimiting examples of heterocycle groupsinclude 1,3-dioxolane, 1,4-dioxane, 1,4-dithiane, 2H-pyran,2-pyrazoline, 4H-pyran, chromanyl, imidazolidinyl, imidazolinyl,indolinyl, isochromanyl, isoindolinyl, morpholine, piperazinyl,piperidine, piperidyl, pyrazolidine, pyrazolidinyl, pyrazolinyl,pyrrolidine, pyrroline, quinuelidine, thiomorpholine, and the like.

[0078] The term “alkylene” refers to a divalent straight or branchedhydrocarbon chain (e.g. ethylene —CH₂CH₂—).

[0079] The term “aryl(C₁-C₈)alkylene” for example includes benzyl,phenethyl, naphthylmethyl and the like.

[0080] The carbon atom content of various hydrocarbon-containingmoieties is indicated by a prefix designating the minimum and maximumnumber of carbon atoms in the moiety, i.e., the prefix C_(i)-C_(j)indicates a moiety of the integer “i” to the integer “j” carbon atoms,inclusive. Thus, for example, (C₁-C₈)alkyl refers to alkyl of one toeight carbon atoms, inclusive.

[0081] The compounds of the present invention are generally namedaccording to the IUPAC or CAS nomenclature system. Abbreviations whichare well known to one of ordinary skill in the art may be used (e.g.,“Ph” for phenyl, “Me” for methyl, “Et” for ethyl, “h” for hour or hoursand “rt” for room temperature).

[0082] Specific and preferred values listed below for radicals,substituents, and ranges, are for illustration only; they do not excludeother defined values or other values within defined ranges for theradicals and substituents.

[0083] Specifically, (C₁-C₈)alkyl can be methyl, ethyl, propyl,isopropyl, butyl, iso-butyl, sec-butyl, pentyl, 3-pentyl, hexyl, orheptyl; (C₁-C₈)alkoxy can be methoxy, ethoxy, propoxy, isopropoxy,butoxy, iso-butoxy, sec-butoxy, pentoxy, 3-pentoxy, hexyloxy,1-methylhexyloxy, or heptyloxy; aryl can be phenyl, indenyl, ornaphthyl; and heteroaryl can be furyl, imidazolyl, triazolyl, triazinyl,oxazoyl, isoxazoyl, thiazolyl, isothiazoyl, pyrazolyl, pyrrolyl,pyrazinyl, tetrazolyl, pyridyl, (or its N-oxide), thienyl, pyrimidinyl(or its N-oxide), indolyl, isoquinolyl (or its N-oxide) or quinolyl (orits N-oxide).

[0084] A specific value for R¹ is hydrogen, —OH, —CH₂OH, —OMe, —OAc,—NH₂, —NHMe, —NMe₂ or —NHAc.

[0085] Another specific value for R¹ is hydrogen, —OH, —OMe, —OAc, —NH₂,—NHMe, —NMe₂ or —NHAc.

[0086] Another specific value for R¹ is hydrogen, —OH, —OMe, or —NH₂.

[0087] Another specific value for R¹ is hydrogen, —OH, or —NH₂—.

[0088] A more specific value for R¹ is hydrogen or —OH.

[0089] A specific value for R¹, R² and the carbon atom to which they areattached is carbonyl (C═O).

[0090] A specific value for R² is hydrogen or (C₁-C₈)alkyl, cyclopropyl,cyclohexyl or benzyl.

[0091] Another specific value for R² is hydrogen, methyl, ethyl orpropyl.

[0092] Another specific value for R² is hydrogen or methyl.

[0093] A more specific value for R² is hydrogen

[0094] A specific value for R³ is hydrogen, OH, OMe, OAc, NH₂, NHMe,NMe₂ or NHAc.

[0095] Another specific value for R³ is hydrogen, OH, OMe, or NH₂.

[0096] Another specific value for R³ is hydrogen, OH, or NH₂.

[0097] A more specific value for R³is hydrogen or OH.

[0098] A specific value for the ring comprising R⁴, R⁵ and the atom towhich they are connected is cyclopentane, cyclohexane, piperidine,dihydro-pyridine, tetrahydro-pyridine, pyridine, piperazine, decaline,tetrahydro-pyrazine, dihydro-pyrazine, pyrazine, dihydro-pyrimidine,tetrahydro-pyrimidine, hexahydro-pyrimidine, pyrazine, imidazole,dihydro-imidazole, imidazolidine, pyrazole, dihydro-pyrazole, and.pyrazolidine.

[0099] A more specific value for the ring comprising R⁴ and R⁵ and theatom to which they are connected is, cyclohexane, piperidine orpiperazine.

[0100] A specific value for R⁶ is (C₁-C₈)alkyl, or substituted(C₁-C₈)alkyl, —OR^(a), —CO₂R^(a), R^(a)C(═O)—, R^(a)C(═O)O—,R^(a)R^(b)N—, R^(a)R^(b)NC(═O)—, or aryl.

[0101] Another specific value for R⁶ is (C₁-C₈)alkyl, —OR^(a),—CO₂R^(a), R^(a)C(═O)—, R^(a)C(═O)O—, R^(a)R^(b)N—, R^(a)R^(b)NC(═O)—,or aryl.

[0102] Another specific value for R⁶ is methyl, ethyl, butyl, OH,OR^(a), —CO₂R^(a), R^(a)C(═O)—, OC(═O)CH₂CH₃, —CONR^(a)R^(b),—NR^(a)R^(b) or phenyl.

[0103] Another specific value for R⁶ is OH, OMe, methyl, ethyl, t-butyl,—CO₂R^(a), —C(═O)NR^(a)R^(b), —OAc, —NH₂, —NHMe, —NMe₂, —NHEt or—N(Et)₂.

[0104] Another specific value for R⁶ is —(CH₂)₁₋₂OR^(a),—(CH₂)₁₋₂C(═O)OR^(a), —(CH₂)₁₋₂OC(═O)R^(a), —(CH₂)₁₋₂C(═O)R^(a),—(CH₂)₁₋₂OCO₂R^(a), —(CH₂)₁₋₂NHR^(a), —(CH₂)₁₋₂NR^(a)R^(b),—(CH₂)₁₋₂OC(═O)NHR^(a), or (CH₂)₁₋₂OC(═O)NR^(a)R^(b).

[0105] Another specific value for R⁶ is —CH₂OH, —CH₂OAc, —CH₂OCH₃,—CH₂C(═O)OCH₃, —CH₂OC(═O)CH₃, —CH₂C(═O)CH₃, —CH₂OCO₂CH₃, —CH₂NH(CH₃), or—(CH₂)₁₋₂N(CH₃)₂.

[0106] Another specific value for R⁶ is methyl, ethyl, t-butyl, phenyl,—CO₂R^(a), —CONR^(a)R^(b), or R^(a)C(═O)—.

[0107] Another specific value for R⁶ is —CH₂OH, —CH₂OAc, —C(═O)OCH₃,—C(═O)CH₃, OCO₂CH₃—OCO₂CH₃, —CH₂NH(CH₃), or —(CH₂)₁₋₂N(CH₃)₂.

[0108] A more specific value for R⁶ is methyl, ethyl,—CO₂R^(a)—CONR^(a)R^(b), or R^(a)C(═O)—.

[0109] A specific number of R⁶ groups substituted on the R⁴R⁵ ring isfrom 1 to about 4.

[0110] A specific value for R^(a) and R^(b) is independently hydrogen,(C₁-C₄)alkyl, aryl or aryl(C₁-C₈)alkylene.

[0111] A specific value for R^(a) and R^(b) is independently hydrogen,methyl, ethyl, phenyl or benzyl.

[0112] A more specific value for R^(a) is (C₁-C₈)alkyl.

[0113] Another specific value for R^(a) is methyl, ethyl, propyl orbutyl.

[0114] A more specific value for R^(a) is methyl, ethyl, i-propyl,i-butyl or tert-butyl.

[0115] Another specific value for R^(a) and R^(b) is a ring

[0116] A specific value for R⁷ is hydrogen, alkyl, aryl oraryl(C₁-C₈)alkylene.

[0117] Another specific value for R⁷ is hydrogen, methyl or ethyl,phenyl or benzyl.

[0118] A more specific value for R⁷ is H, or methyl.

[0119] A specific value for —N(R⁷)₂ is amino, methylamino,dimethylamino, ethylamino, pentylamino, diphenylethylamino,pyridylmethylamino, diethylamino or benzylamino.

[0120] A specific value for —N(R⁷)₂ is amino, methylamino,dimethylamino, ethylamino, diethylamino diphenylethylamino, pentylaminoor benzylamino.

[0121] A specific value for N(R⁷)₂ is amino, or methylamino.

[0122] A specific value for X is —CH₂OR^(a), —CO₂R^(a), —OC(O)R^(a),—CH₂OC(O)R^(a), —C(O)NR^(a)R^(b).

[0123] Another specific value for X is —CH₂OR^(a) or —C(O)NR^(a)R^(b).

[0124] A more specific value for X is —CH₂OH or —C(O)NHCH₂CH₃.

[0125] A specific value for m is 0, 1, or 2.

[0126] A more specific value for m is 0, or 1.

[0127] Specific examples of rings comprising R⁴, R⁵ and the atom towhich they are connected include:

[0128] where q is from 0 to 14 and R^(d) is hydrogen, provided that whenq is zero then R^(d) is not hydrogen.

[0129] More specific examples of rings comprising R⁴, R⁵ and the atom towhich they are connected include:

[0130] A specific value for the ring comprising —C(R³)R⁴R⁵ is 2-methylcyclohexane, 2,2-dimethylcyclohexane, 2-phenylcyclohexane,2-ethylcyclohexane, 2,2-diethylcyclohexane, 2-tert-butyl cyclohexane,3-methyl cyclohexane, 3,3-dimethylcyclohexane, 4-methyl cyclohexane,4-ethylcyclohexane, 4-phenyl cyclohexane, 4-tert-butyl cyclohexane,4-carboxymethyl cyclohexane, 4-carboxyethyl cyclohexane,3,3,5,5-tetramethyl cyclohexane, 2,4-dimethyl cyclopentane.4-cyclohexanecarboxyic acid, 4-cyclohexanecarboxyic acid esters, or4-methyloxyalkanoyl-cyclohexane.

[0131] A specific value for the ring comprising —C(R³)R⁴R⁵ is4-piperidine, 4-piperidene-1-carboxylic acid, 4-piperidine-1-carboxylicacid methyl ester, 4-piperidine-1-carboxylic acid ethyl ester,4-piperidine-1-carboxylic acid propyl ester, 4-piperidine-1 -carboxylicacid tert-butyl ester, 1-piperidine, 1-piperidine-4-carboxylic acidmethyl ester, 1-piperidine-4-carboxylic acid ethyl ester,1-piperidine-4-carboxylic acid propyl ester, 1-piperidine-4-caboxylicacid tert-butyl ester, 1-piperidine-4-carboxylic acid methyl ester,3-piperidine, 3-piperidene-1-carboxylic acid, 3-piperidine-1-carboxylicacid methyl ester, 3-piperidine-1-carboxylic acid tert-butyl ester,1,4-piperazine, 4-piperazine-1-carboxylic acid,4-piperazine-1-carboxylic acid methyl ester, 4-piperazine-1-carboxylicacid ethyl ester, 4-piperazine-1-carboxylic acid propyl ester,4-piperazine-1-carboxylic acid tert-butylester, 1,3-piperazine,3-piperazine-1-carboxylic acid, 3-piperazine-1-carboxylic acid methylester, 3-piperazine-1-carboxylic acid ethyl ester,3-piperazine-1-carboxylic acid propyl ester, 3-piperidine-1-carboxylicacid tert-butylester, 1-piperidine-3-carboxylic acid methyl ester,1-piperidine-3-carboxylic acid ethyl ester, 1-piperidine-3-carboxylicacid propyl ester or 1-piperidine-3-caboxylic acid tert-butyl ester.

[0132] A specific value for the ring comprising R⁴ and R⁵ is 2-methylcyclohexane, 2,2-dimethylcyclohexane, 2-phenyl cyclohexane,2-ethylcyclohexane, 2,2-diethylcyclohexane, 2-tert-butyl cyclohexane,3-methyl cyclohexane, 3,3-dimethylcyclohexane, 4-methyl cyclohexane,4-ethylcyclohexane, 4-phenyl cyclohexane, 4-tert-butyl cyclohexane,4-carboxymethyl cyclohexane, 4-carboxyethyl cyclohexane,3,3,5,5-tetramethyl cyclohexane, 2,4-dimethyl cyclopentane,4-piperidine-1-carboxylic acid methyl ester, 4-piperidine-1-carboxylicacid tert-butyl ester 4-piperidine, 4-piperazine-1-carboxylic acidmethyl ester, 4-piperidine-1-carboxylic acid tert-butylester,1-piperidine-4-carboxylic acid methyl ester, 1-piperidine-4-caboxylicacid tert-butyl ester, tert-butylester, 1-piperidine-4-carboxylic acidmethyl ester, or 1-piperidine-4-caboxylic acid tert-butyl ester,3-piperidine-1-carboxylic acid methyl ester, 3-piperidine-1-carboxylicacid tert-butyl ester, 3-piperidine, 3-piperazine-1-carboxylic acidmethyl ester, 3-piperidine-1-carboxylic acid tert-butylester,1-piperidine-3-carboxylic acid methyl ester, 1-piperidine-3-caboxylicacid tert-butyl ester

[0133] In another embodiment the invention includes a compound havingthe general formula (I):

[0134] (I) wherein

[0135] Z is CR³R⁴R⁵ or NR⁴R⁵;

[0136] each R¹ is independently hydrogen, halo, —OR^(a), —SR^(a),(C₁-C₈)alkyl, cyano, nitro, trifluoromethyl, trifluoromethoxy,(C₃-C₈)cycloalkyl, heterocycle, hetrocycle(C₁-C₈)alkylene-, aryl,aryl(C₁-C₈)alkylene-, heteroaryl, heteroaryl(C₁-C₈)alkylene-, —CO₂R^(a),R^(a)C(═O)O—, R^(a)C(═O)—, —OCO₂R^(a), R^(a)R^(b)NC(═O)O—,R^(b)OC(═O)N(R^(a))—, R^(a)R^(b)N—, R^(a)R^(b)NC(═O)—,R^(a)C(═O)N(R^(b))—, R^(a)R^(b)NC(═O)N(R^(b))—,R^(a)R^(b)NC(═S)N(R^(b))—, —OPO₃R^(a), R^(a)OC(═S)—, R^(a)C(═S)—,—SSR^(a), R^(a)S(═O)—, —N═NR^(a), or —OPO₂R^(a);

[0137] each R² independently hydrogen, (C₁-C₈)alkyl, (C3-C₈)cycloalkyl,heterocycle, heterocycle(C₁-C₈)alkylene-, aryl, aryl(C₁-C₈)alkylene-,heteroaryl, or heteroaryl(C₁-C₈)alkylene-; or

[0138] R¹ and R² and the atom to which they are attached can be C═O orC═NR^(c).

[0139] R⁴ and R⁵ together with the atoms to which they are attached canform a saturated or unsaturated, mono-, bicyclic- or aromatic ringhaving 3, 4, 5, 6, 7 or 8 ring atoms optionally comprising 1, 2, 3, or 4heteroatoms selected from oxy (—O—), thio (—S—), sulfinyl (—SO—),sulfonyl (—S(O)₂—) or amine (—NR^(a)—) in the ring;

[0140] wherein any ring comprising R⁴ and R⁵ is substituted with from 1to 14 R⁶ groups; wherein each R⁶ is independently halo, —OR^(a),—SR^(a), (C₁-C₈)alkyl, cyano, nitro, trifluoromethyl, trifluoromethoxy,(C₃-C₈)cycloalkyl, (C₃-C₈)bicycloalkyl, heterocycle orhetrocycle(C₁-C₈)alkylene-, aryl, aryl(C₁-C₈)alylene-, heteroaryl,heteroaryl(C₁-C₈)alkylene-, —CO₂R^(a), R^(a)C(═O)O—, R^(a)C(═O)—,—OCO₂R^(a), R^(a)R^(b)NC(═O)O—, R^(b)OC(═O)N(R^(a))—, R^(a)R^(b)N—,R^(a)R^(b)NC(═O)—, R^(a)C(═O)N(R^(b))—, R^(a)R^(b)NC(═O)N(R^(b))—,R^(a)R^(b)NC(═S)N(R^(b))—, —OPO₃R^(a), R^(a)OC(═S—, R^(a)C(═S)—,—SSR^(a), R^(a)S(═O)—, —NNR¹ or —OPO₂R^(a);

[0141] R³ is hydrogen, halo, —OR^(a), —SR^(a), C₁₋₈alkyl, cyano, nitro,trifluoromethyl, trifluoromethoxy, (C₃-C₈)cycloalkyl, heterocycle orhetrocycle(C₁-C₈)alkylene-, aryl, aryl(C₁-C₈)alkylene-, heteroaryl,heteroaryl(C₁-C₈)alkylene-, —CO₂R^(a), R^(a)C(═O)O—, R^(a)C(O)—,—OCO₂R^(a), R^(a)R^(b)NC(═O)O—, R^(b)OC(═O)N(R^(a))—, R^(a)R^(b)N—,R^(a)R^(b)NC(═O)—, R^(a)C(═O)N(R^(b))—, R^(a)R^(b)NC(═O)N(R^(b))—,R^(a)R^(b)NC(═S)N(R^(b))—, —OPO₃R^(a), R^(a)OC(═S)—, R^(a)C(═S)—,—SSR^(a), R^(a)S(═O)—, —N═NR^(a), —OPO₂R^(a); or if the ring formed fromCR⁴R⁵ is aryl or hereroaryl or partially unsaturated then R³ can beabsent;

[0142] each R⁷ is independently hydrogen, (C₁-C₈)alkyl,(C₃-C₈)cycloalkyl, aryl or aryl(C₁-C₈)alkylene;

[0143] X is —CH₂OR^(a), —CO₂R^(a), —OC(O)R^(a), —CH₂OC(O)R^(a),C(O)NR^(a)R^(b), —CH₂SR^(a), —C(S)OR^(a), —OC(S)R^(a), —CH₂OC(S)R^(a) orC(S)NR^(a)R^(b) or —CH₂N(R^(a))(R^(b));

[0144] wherein any of R¹, R², R³ and R⁶ is optionally substituted with(C₁-C₈)alkyl, aryl, heteroaryl, heterocycle, aryloxy, (C₃-C₈)cycloalkyl,hydroxy, nitro, halo, cyano, (C₁-C₈)alkoxy, (C₁-C₈)alkanoyl,(C₁-C₈)alkoxycarbonyl, (C₁-C₈)alkanoyloxy, R^(a)S(O)_(p)—,R^(a)R^(b)NS(O)—, R^(a)R^(b)NS(O)₂—, R^(a)R^(b)N—, or R^(a)R^(b)NC(═O)—;

[0145] wherein any (C₁-C₈)alkyl, (C₃-C₈)cycloalkyl, (C₃-C₈)bicycloalkyl,(C₁-C₈)alkoxy, (C₁-C₈)alkanoyl, (C₁-C₈)alkylene, or heterocycle, isoptionally partially unsaturated;

[0146] R^(a) and R^(b) are each independently hydrogen, (C₁-C₈)alkyl, or(C₁-C₈)alkyl substituted with 1-3 (C₁-C₈)alkoxy, (C₃-C₈)cycloalkyl,(C₁-C₈)alkylthio, amino acid, aryl, aryl(C₁-C₈)alkylene, heteroaryl, orheteroaryl(C₁-C₈)alkylene;; or R^(a) and R^(b), together with thenitrogen to which they are attached, form a pyrrolidino, piperidino,morpholino, or thiomorpholino ring; and R^(c) is hydrogen or C₁₋₆ alkyl;

[0147] m is 0 to about 8 and p is 0 to 2; provided that when m is 0 orall R¹ and R² groups present are hydrogen then R³ is not hydrogen;provided that m is at least 1 when Z is NR⁴R⁵; or a pharmaceuticallyacceptable salt thereof.

[0148] Specific compounds of formula (I) are those wherein each R⁷ is H,X is ethylaminocarbonyl and

[0149] R¹ is hydroxy, R² is hydrogen, and Z is 4-carboxycyclohexyl,wherein R^(a) is hydrogen, 4; Z is 4-methoxycarbonylcyclohexylmethyl,R^(a) is methyl, 5; R¹ and R² together are oxo, Z is a4-carbonylcyclohexyl group, wherein R^(a) is methyl, methoxy, ethyl,ethoxy, propyl, isopropoxy, -isobutyl, tert-butyl, amine, methylamine ordimethylamine, 6.

[0150] Another group of specific compounds of formula (I) are thosewherein each R⁷ is H, X is ethylaminocarbonyl,

[0151] R¹ is hydroxy, R² is hydrogen, and Z is a substituted4-(methyleneoxy-carbonyl)cyclohexyl group, wherein R^(a) is methyl,ethyl, propyl, tert-butyl, methoxy, ethoxy, methylamine ordimethylamine, 7; or R¹ and R² together are oxo, and Z is a substituted-(methyleneoxycarbonyl)cyclohexyl group, wherein R^(a) is methyl, ethyl,propyl, tert-butyl, methoxy, ethoxy, methylamine or dimethylamine, 8.

[0152] Another group of specific compounds of formula (1) are thosewherein each R⁷ is H, X is ethylaminocarbonyl, and

[0153] R¹ and R² are each hydrogen, and Z is a 1-piperidyl-4-carboxylicacid or ester group, wherein R^(a) is hydrogen, methyl, ethyl, propyl,isopropyl, or t-butyl, 9; R¹ and R² together are oxo, and Z is a1-piperidyl-4-carboxylic acid or ester group, wherein R^(a) is hydrogen,methyl, ethyl, propyl, isopropyl, or t-butyl, 10; R¹ and R² are eachhydrogen and Z is a 4-(methyleneoxy-carbonyl)piperidin-4-yl groupwherein R^(a) is methyl, ethyl, propyl or t-butyl, amine, methylamine,dimethylamine, 11; or R¹ and R² together are oxo, and Z is a4-(methyleneoxycarbonyl)piperidin-4-yl wherein R^(a) is methyl, ethyl,propyl or t-butyl, amine, methylamine, dimethylamine, 12; R¹ and R² areeach hydrogen and Z is a 4-(methyleneoxycarbonyl)piperidin-4-yl-oxywherein R^(a) is hydrogen, methyl, ethyl, propyl isopropyl, isobutyl, ort-butyl, 13 or R¹ and R² together are oxo, Z is a4-(methyleneoxycarbonyl)piperidin-4-yl-oxy wherein R^(a) is hydrogen,methyl, ethyl, propyl, isopropyl, isobutyl, or t-butyl, 14.

[0154] Another group of specific compounds of formula (I) are thosewherein each R⁷ is H, X is ethylaminocarbonyl,

[0155] R¹ and R² are each hydrogen, and Z is a 4-piperidyl-1-carboxylicacid or ester group, wherein R^(a) is methyl, ethyl, propyl, isopropyl,isobutyl, or t-butyl, 15, R¹ is hydroxy, R² is hydrogen, and Z is a4-piperidyl-1-carboxylic acid or ester group, wherein R^(a) is metho,ethyl, propyl, isopropyl, isobutyl, or t-butyl, 16; or R¹ and R²together are oxo, and Z is a 4-piperidyl-1-carboxylic acid or estergroup, wherein R¹ is methyl, ethyl, propyl, isopropyl, isobutyl, ort-butyl, 17.

[0156] Another group of specific compounds of formula (I) are thosewherein each R⁷ is H, X is ethylaminocarbonyl,

[0157] R¹ and R² are each hydrogen, Z is a 4-piperazine-1-carboxylicacid or ester group wherein R^(a) is methyl, ethyl, isopropyl, isobutyl,or t-butyl, 18; or R¹ and R² together are oxo, Z is a4-piperazine-1-carboxylic acid or ester group wherein R^(a) is methyl,ethyl, isopropyl, isobutyl, or t-butyl, 19.

[0158] Additional compounds of the invention are depicted in tables 1,2, 3, 4, 5, 6 and 7 below: TABLE 1

Compound R R¹ R² R⁶ ATL2037 NECA H H CH₂OH MP9056 NECA OH H CH₂OHATL146a NECA H H CO₂H MP9057 NECA OH H CO₂H ATL146e NECA H H CO₂MeMP9058 NECA OH H CO₂Me JR2145 CH₂OH H H CO₂Me MP9059 CH₂OH OH H CO₂MeATL193 NECA H H CH₂OAc MP9060 NECA OH H CH₂Oac JR2147 CH₂OH H H CH₂OacMP9061 CH₂OH OH H CH₂Oac JR3023 NECA H H CH₂N(CH₃)₂ MP9062 NECA OH HCH₂N(CH₃)₂ JR3021 NECA H H COOCH₂CH₂NHBoc MP9063 NECA OH HCOOCH₂CH₂NHBoc JR3033 NECA H H COOCH₂CH₂NH₂ MP9064 NECA OH HCOOCH₂CH₂NH₂ JR3037 NECA H H CONHCH₂CH₃ MP9065 NECA OH H CONHCH₂CH₃JR3055 NECA H H CONH₂ MP9072 NECA OH H CONH₂ JR3065 NECA H H CONHMeMP9066 NECA OH H CONHMe JR3067B NECA H H Me, cis CO₂Me MP9067 NECA OH HMe, cis CO₂Me JR3067A NECA H H Me, trans CO₂Me MP9068 NECA OH H Me,trans CO₂Me JR3087 NECA H H CH₂CH₃ MP9069 NECA OH H CH₂CH₃ JR3159A NECAOH H H JR3159B NECA OH H H JR3119 NECA H H COCH₃ MP9070 NECA OH H COCH₃JR3121 NECA H H CHCH₃(OH) MP9071 NECA OH H CHCH₃(OH) JR3139 NECA OHC₆H₁₁ H

[0159] TABLE 2

Compound R¹ R² R⁶ JR3261 H H H JR3259 H H CO₂tBu JR3269 H H CO₂Et JR4011H H CO₂iBu JR4009 H H CO₂iPr JR4007 H H COMe JR4051 H H COC(CH₃)₃ JR4047H H COCH₂(CH₃)₃ MP9047 H H COCH₃ MP9048 H H C(O)N(CH₃)₂ MP9049 H HC(O)N(CH₃)Et MP9050 H H C(O)N(CH₃)iPr MP9051 H H C(O)N(CH₃)iBu MP9052 HH C(O)NH(CH₃) MP9053 H H C(O)NH(Et) MP9054 H H C(O)NH(iPr) MP9055 H HC(O)NH(iBu) TX3261 OH H H TX3259 OH H CO₂tBu TX3269 OH H CO₂Et TX4011 OHH CO₂iBu TX4009 OH H CO₂iPr TX4007 OH H COMe TX4051 OH H COC(CH₃)₃TX4047 OH H COCH₂(CH₃)₃ TX9047 OH H COCH₃ TX9048 OH H C(O)N(CH₃)₂ TX9049OH H C(O)N(CH₃)Et TX9050 OH H C(O)N(CH₃)iPr TX9051 OH H C(O)N(CH₃)iBuTX9052 OH H C(O)NH(CH₃) TX9053 OH H C(O)NH(Et) TX9054 OH H C(O)NH(iPr)TX9055 OH H C(O)NH(iBu)

[0160] TABLE 3

Compound n R³ R⁶ JR3135 1 OH H JR3089 2 OH H JR3205 2 NH₂ H JR3177A 2 OH2-CH₃ JR3177B 2 OH 2-CH₃ JR3181A 2 OH 2-CH₃ JR3181B 2 OH 2-CH₃ JR3227 2OH 2-C(CH₃)₃ JR9876 2 OH 2-C₆H₅ JR3179 2 OH 3-CH₃ JR3221 2 OH(R)3-CH₃(R) JR3223 2 OH(S) 3-CH₃(R) MP9041 2 OH(R) 3-CH₃(S) MP9042 2 OH(S)3-CH₃(S) JR3201B 2 OH 3-(CH₃)₂ MP9043 2 OH(R) 3-CH₂CH₃(R) MP9044 2 OH(S)3-CH₂CH₃(R) MP9045 2 OH(R) 3-CH₂CH₃(S) MP9046 2 OH(S) 3-CH₂CH₃(S) JR31632 OH 3-(CH₃)₂, 5-(CH₃)₂ JR9875 2 OH 4-CH₃ JR3149 2 OH 4-C₂H₅ JR3203 2 OH4-C(CH₃)₃ JR3161 2 OH 4-C₆H₅

[0161] TABLE 4

Compound R¹ R² R⁶ JR3213 H H CO₂Et JR3281 H H CO₂tBu JR3289 H H H JR4025H H cyclohexyl JR4053 H H COMe JR4049 H H CO₂iBu JR3283 H H2-Pyrimidinyl MP9029 H H COMe MP9030 H H COC(CH₃)₃ MP9031 H HCOCH₂(CH₃)₃ MP9032 H H COCH₃ MP9033 H H C(O)N(CH₃)₂ MP9034 H HC(O)N(CH₃)Et MP9035 H H C(O)N(CH₃)iPr MP9036 H H C(O)N(CH₃)iBu MP9037 HH C(O)NH(CH₃) MP9038 H H C(O)NH(Et) MP9039 H H C(O)NH(iPr) MP9040 H HC(O)NH(iBu)

[0162] TABLE 5

Compound R R¹ R² R⁶ MP9021 NECA H H CH₂OH MP9022 NECA H H CO₂H JR3251NECA H H CO₂Me JR3279 NECA H H CO₂Et MP9027 CH₂OH H H CO₂Me MP9028 NECAH H CO₂MeCH₂OAc MP9015 CH₂OH H H CH₂OAc MP9016 NECA H H CH₂N(CH₃)₂MP9017 NECA H H COOCH₂CH₂NHBoc MP9018 NECA H H COOCH₂CH₂NH₂ MP9019 NECAH H CONHCH₂CH₃ MP9020 NECA H H CONH₂ MP9023 NECA H H CONHMe MP9024 NECAH H CH₂CH₃ MP9025 NECA H H COCH₃ MP9026 NECA H H CHCH₃(OH)

[0163] TABLE 6

Compound R R¹ R² R⁶ MP9001 NECA H H CH₂OH MP9002 NECA H H CO₂H JR3253NECA H H CO₂Me MP9003 CH₂OH H H CO₂Me MP9004 NECA H H CH₂OAc MP9005CH₂OH H H CH₂OAc MP9006 NECA H H CH₂N(CH₃)₂ MP9007 NECA H HCOOCH₂CH₂NHBoc MP9008 NECA H H COOCH₂CH₂NH₂ MP9009 NECA H H CONHCH₂CH₃MP9010 NECA H H CONH₂ MP9011 NECA H H CONHMe MP9012 NECA H H CH₂CH₃MP9013 NECA H H COCH₃ MP9014 NECA H H CHCH₃(OH)

[0164] TABLE 7

Compound R Y Y′ R6 RJ1111 NECA CH CH CO₂Me RJ1112 NECA CH N CO₂Me RJ1113NECA N CH CO₂Me RJ1114 NECA N N CO₂Me RJ1115 NECA CH CH CH₂OH RJ1116NECA CH N CH₂OH RJ1117 NECA N CH CH₂OH RJ1118 NECA N N CH₂OH RJ1119 NECACH CH CO₂H RJ1120 NECA CH N CO₂H RJ1121 NECA N CH CO₂H RJ1122 NECA N NCO₂H RJ1123 NECA CH CH CH₂OAc RJ1124 NECA CH N CH₂OAc RJ1125 NECA N CHCH₂OAc RJ1126 NECA N N CH₂OAc RJ1127 NECA CH CH CONH₂ RJ1128 NECA CH NCONH₂ RJ1129 NECA N CH CONH₂ RJ1130 NECA N N CONH₂ RJ1131 NECA CH CHCONHMe RJ1132 NECA CH N CONHMe RJ1133 NECA N CH CONHMe RJ1134 NECA N NCONHMe RJ1135 NECA CH CH CO₂tBu RJ1136 NECA CH N CO₂tBu RJ1137 NECA N CHCO₂tBu RJ1138 NECA N N CO₂tBu RJ1139 NECA CH CH CO₂Et RJ1140 NECA CH NCO₂Et RJ1141 NECA N CH CO₂Et RJ1142 NECA N N CO₂Et RJ1143 NECA CH CHCO₂iBu RJ1144 NECA CH N CO₂iBu RJ1145 NECA N CH CO₂iBu RJ1146 NECA N NCO₂iBu RJ1147 NECA CH CH CO₂iPr RJ1148 NECA CH N CO₂iPr RJ1149 NECA N CHCO₂iPr RJ1150 NECA N N CO₂iPr RJ1151 NECA CH CH COMe RJ1152 NECA CH NCOMe RJ1153 NECA N CH COMe RJ1154 NECA N N COMe RJ1155 NECA CH CHCOC(CH₃)₃ RJ1156 NECA CH N COC(CH₃)₃ RJ1157 NECA N CH COC(CH₃)₃ RJ1158NECA N N COC(CH₃)₃ RJ1159 NECA CH CH COCH₂(CH₃)₃ RJ1160 NECA CH NCOCH₂(CH₃)₃ RJ1161 NECA N CH COCH₂(CH₃)₃ RJ1162 NECA N N COCH₂(CH₃)₃RJ1163 NECA CH CH C(O)N(CH₃)₂ RJ1164 NECA CH N C(O)N(CH₃)₂ RJ1165 NECA NCH C(O)N(CH₃)₂ RJ1166 NECA N N C(O)N(CH₃)₂ RJ1167 NECA CH CHC(O)N(CH₃)Et RJ1168 NECA CH N C(O)N(CH₃)Et RJ1169 NECA N CH C(O)N(CH₃)EtRJ1170 NECA N N C(O)N(CH₃)Et RJ1171 NECA CH CH C(O)N(CH₃)iPr RJ1172 NECACH N C(O)N(CH₃)iPr RJ1173 NECA N CH C(O)N(CH₃)iPr RJ1174 NECA N NC(O)N(CH₃)iPr RJ1175 NECA CH CH C(O)N(CH₃)iBu RJ1176 NECA CH NC(O)N(CH₃)iBu RJ1177 NECA N CH C(O)N(CH₃)iBu RJ1178 NECA N NC(O)N(CH₃)iBu RJ1179 NECA CH CH C(O)NH(Et) RJ1180 NECA CH N C(O)NH(Et)RJ1181 NECA N CH C(O)NH(Et) RJ1182 NECA N N C(O)NH(Et) RJ1183 NECA CH CHC(O)NH(iPr) RJ1184 NECA CH N C(O)NH(iPr) RJ1185 NECA N CH C(O)NH(iPr)RJ1186 NECA N N C(O)NH(iPr) RJ1187 NECA CH CH C(O)NH(iBu) RJ1188 NECA CHN C(O)NH(iBu) RJ1189 NECA N CH C(O)NH(iBu) RJ1190 NECA N N C(O)NH(iBu)RJ1191 NECA CH CH CH₂OCOCH₃ RJ1192 NECA N CH CH₂OCOCH₃ RJ1193 NECA CH CHCH₂OCOEt RJ1194 NECA N CH CH₂OCOEt RJ1195 NECA CH CH CH₂OCOiPr RJ1196NECA N CH CH₂OCOiPr RJ1197 NECA CH CH CH₂OCOiBu RJ1198 NECA N CHCH₂OCOiBu

[0165] The following abbreviations have been used herein: 2-Aas2-alkynyladenosines; ¹²⁵I-ABA N⁶-(4-amino-3-¹²⁵iodo-benzyl)adenosineAPCI Atmospheric pressure chemical ionization ATL146e4-{3-[6-Amino-9-(5-ethylcarbamoyl-3,4-dihydroxy-tetrahydro-furan-2-yl)-9H-purin-2-yl]-prop-2-ynyl}cyclo-hexanecarboxylic acid methyl ester; CCPA2-chloro-N⁶-cyclopentyladenosine; CGS216802-[14-(2-carboxyethyl)phenethylamino]-5′-N-ethyl- carboxamidoadenosine;Cl-IB-MECA N⁶-3-iodo-2-chlorobenzyladenosine-5′-N- methyluronamide; CPAN⁶-cyclopentyladenosine DMF dimethylformamide DMSO dimethylsulfoxideDMSO-d₆ deuterated dimethylsulfoxide EtOAc ethyl acetate eq equivalentGPCR G protein coupled receptor; hA_(2A)AR, Recombinant human A_(2A)adenosine receptor; IADO 2-Iodoadenosine ¹²⁵I-APE,2-[2-(4-amino-3-[¹²⁵I]iodophenyl)ethylamino] adenosine; NECA,5′-N-ethylcarboxamidoadenosine; IB-MECAN⁶-3-iodobenzyladenosine-5′-N-methyluronamide; 2-Iodoadenosine5-(6-amino-2-iodo-purin-9-yl)-3,4- dihydroxytetrahydro-furan-2carboxylicacid ethylamide HPLC high-performance liquid chromatography HRMShigh-resolution mass spectrometry ¹²⁵I-ZM241385,¹²⁵I-4-(2[7-amino-2-[2-furyl][1,2,4]triazolo[2,3-a][1,3,5]-triazin-5-yl-amino]ethyl)phenol; INECA2-iodo-N-ethylcarboxamidoadenosine LC/MS liquid chromatography/massspectrometry m.p. melting point MHz megahertz MRS 1220,N-(9-chloro-2-furan-2-yl-[1,2,4]triazolo[1,5-c]-quinazolin-5-yl)-2-phenylacetamide; MS mass spectrometry NECAN-ethylcarboxamidoadenosine NMR nuclear magnetic resonance RP-HPLCreversephase high-performance liquid chromatography TBAFtetrabutylammonium fluoride TBS tert-butyldimethylsilyl TBDMSCltert-butyldimethylsilylchloride TEA triethylamine TFA trifluoroaceticacid THF tetrahydrofuan TLC thin layer chromatography p-TSOHpara-toluenesulfonic acid XAC8-(4-((2-a-minoethyl)aminocarbonyl-methyloxy)-phenyl)-1-3-dipropylxanthine;

[0166] Compounds of the invention can generally be prepared asillustrated in Schemes 1A and 1B below. Starting materials can beprepared by procedures described in these schemes, procedures describedin the General methods below or by procedures that would be well knownto one of ordinary skill in organic chemistry. The variables used inSchemes 1A and Scheme 1B are as defined herein or as in the claims.

[0167] The preparation of alkynyl cycloalkanols is illustrated in Scheme1A. A solution of an appropriate cycloalkanone (where j is from 0-5) isprepared in a solvent such as THF. A solution of a suitableethynylmagnesium halide compound in a solvent is added to thecycloalkanone. After additioin, the solution is allowed to stir at about20° C. for about 20 hours. The reaction is monitored via TLC until thestarting material is consumed. The reaction is quenched with water,filtered over a plug of sand and silica, washed with a solvent, such asEtOAc, and evaporated to provide the product. Typically, two productsare formed, the isomers formed by the axial/equatorial addition of thealkyne (where m is as defined above, and the sum of m1 and m2 is from 0to about 7) to the ketone. The compounds are purified via flashchromatography using EtOAc/Hexanes to provide the product.

[0168] The preparation of 2-alkynyladenosines is illustrated in Scheme1B. A flame-dried round bottom under nitrogen is charged with5-(6-Amino-2-iodo-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide (NECA 2-Todoadenosine) and a solvent such as DMF. Theappropriate alkyne, wherein R is a —(CR₁R₂)_(m)Z group, is dissolved inacetonitrile followed by TEA, 5 mole % Pd(PPh3)4, and CuI. All solventsare thoroughly degassed.

[0169] The solution is allowed to stir for about 24 hours at roomtemperature, and monitored until complete by HPLC. If the reaction isnot complete after this time, additional catalyst, CuI, and TEA areadded. After the reaction is complete, the solvents are removed underhigh-vacuum and the residue taken up in a small amount of DMF. Thisproduct is isolated using preparative silica TLC. The product ispurified by RP-HPLC.

[0170] Examples of pharmaceutically acceptable salts are organic acidaddition salts formed with acids which form a physiological acceptableanion, for example, tosylate, methanesulfonate, malate, acetate,citrate, malonate, tartarate, succinate, benzoate, ascorbate,α-ketoglutarate, and α-glycerophosphate. Suitable inorganic salts mayalso be formed, including hydrochloride, sulfate, nitrate, bicarbonate,and carbonate salts.

[0171] Pharmaceutically acceptable salts may be obtained using standardprocedures well known in the art, for example by reacting a sufficientlybasic compound such as an amine with a suitable acid affording aphysiologically acceptable anion. Alkali metal (for example, sodium,potassium or lithium) or alkaline earth metal (for example calcium)salts of carboxylic acids can also be made.

[0172] The compounds of formula I can be formulated as pharmaceuticalcompositions and administered to a mammalian host, such as a humanpatient in a variety of forms adapted to the chosen route ofadministration, i.e., orally or parenterally, by intravenous,intramuscular, topical or subcutaneous routes.

[0173] Thus, the present compounds may be systemically administered,e.g., orally, in combination with a pharmaceutically acceptable vehiclesuch as an inert diluent or an assimilable edible carrier. They may beenclosed in hard or soft shell gelatin capsules, may be compressed intotablets, or may be incorporated directly with the food of the patient'sdiet. For oral therapeutic administration, the active compound may becombined with one or more excipients and used in the form of ingestibletablets, buccal tablets, troches, capsules, elixirs, suspensions,syrups, wafers, and the like. Such compositions and preparations shouldcontain at least 0.1% of active compound. The percentage of thecompositions and preparations may, of course, be varied and mayconveniently be between about 2 to about 60% of the weight of a givenunit dosage form. The amount of active compound in such therapeuticallyuseful compositions is such that an effective dosage level will beobtained.

[0174] The tablets, troches, pills, capsules, and the like may alsocontain the following: binders such as gum tragacanth, acacia, cornstarch or gelatin; excipients such as dicalcium phosphate; adisintegrating agent such as corn starch, potato starch, alginic acidand the like; a lubricant such as magnesium stearate; and a sweeteningagent such as sucrose, fructose, lactose or aspartame or a flavoringagent such as peppermint, oil of wintergreen, or cherry flavoring may beadded. When the unit dosage form is a capsule, it may contain, inaddition to materials of the above type, a liquid carrier, such as avegetable oil or a polyethylene glycol. Various other materials may bepresent as coatings or to otherwise modify the physical form of thesolid unit dosage form. For instance, tablets, pills, or capsules may becoated with gelatin, wax, shellac or sugar and the like. A syrup orelixir may contain the active compound, sucrose or fructose as asweetening agent, methyl and propylparabens as preservatives, a dye andflavoring such as cherry or orange flavor. Of course, any material usedin preparing any unit dosage form should be pharmaceutically acceptableand substantially non-toxic in the amounts employed. In addition, theactive compound may be incorporated into sustained-release preparationsand devices.

[0175] The active compound may also be administered intravenously orintraperitoneally by infusion or injection. Solutions of the activecompound or its salts can be prepared in water, optionally mixed with anontoxic surfactant. Dispersions can also be prepared in glycerol,liquid polyethylene glycols, triacetin, and mixtures thereof and inoils. Under ordinary conditions of storage and use, these preparationscontain a preservative to prevent the growth of microorganisms.

[0176] The pharmaceutical dosage forms suitable for injection orinfusion can include sterile aqueous solutions or dispersions or sterilepowders comprising the active ingredient which are adapted for theextemporaneous preparation of sterile injectable or infusible solutionsor dispersions, optionally encapsulated in liposomes. In all cases, theultimate dosage form must be sterile, fluid and stable under theconditions of manufacture and storage. The liquid carrier or vehicle canbe a solvent or liquid dispersion medium comprising, for example, water,ethanol, a polyol (for example, glycerol, propylene glycol, liquidpolyethylene glycols, and the like), vegetable oils, nontoxic glycerylesters, and suitable mixtures thereof. The proper fluidity can bemaintained, for example, by the formation of liposomes, by themaintenance of the required particle size in the case of dispersions orby the use of surfactants. The prevention of the action ofmicroorganisms can be brought about by various antibacterial andantifungal agents, for example, parabens, chlorobutanol, phenol, sorbicacid, thimerosal, and the like. In many cases, it will be preferable toinclude isotonic agents, for example, sugars, buffers or sodiumchloride. Prolonged absorption of the injectable compositions can bebrought about by the use in the compositions of agents delayingabsorption, for example, aluminum monostearate and gelatin.

[0177] Sterile injectable solutions are prepared by incorporating theactive compound in the required amount in the appropriate solvent withvarious of the other ingredients enumerated above, as required, followedby filter sterilization. In the case of sterile powders for thepreparation of sterile injectable solutions, the preferred methods ofpreparation are vacuum drying and the freeze drying techniques, whichyield a powder of the active ingredient plus any additional desiredingredient present in the previously sterile-filtered solutions.

[0178] For topical administration, the present compounds may be appliedin pure form, i.e., when they are liquids. However, it will generally bedesirable to administer them to the skin as compositions orformulations, in combination with a dermatologically acceptable carrier,which may be a solid, a liquid or in a dermatological patch.

[0179] Useful solid carriers include finely divided solids such as talc,clay, microcrystalline cellulose, silica, alumina and the like. Usefulliquid carriers include water, alcohols or glycols orwater-alcohol/glycol blends, in which the present compounds can bedissolved or dispersed at effective levels, optionally with the aid ofnon-toxic surfactants. Adjuvants such as fragrances and additionalantimicrobial agents can be added to optimize the properties for a givenuse. The resultant liquid compositions can be applied from absorbentpads, used to impregnate bandages and other dressings, or sprayed ontothe affected area using pump-type or aerosol sprayers.

[0180] Thickeners such as synthetic polymers, fatty acids, fatty acidsalts and esters, fatty alcohols, modified celluloses or modifiedmineral materials can also be employed with liquid carriers to formspreadable pastes, gels, ointments, soaps, and the like, for applicationdirectly to the skin of the user.

[0181] Examples of useful dermatological compositions, which can be usedto deliver the compounds of formula I to the skin are disclosed inJacquet et al. (U.S. Pat. No. 4,608,392), Geria (U.S. Pat. No.4,992,478), Smith et al. (U.S. Pat. No. 4,559,157) and Wortzman (U.S.Pat. No. 4,820,508).

[0182] Useful dosages of the compounds of formula I can be determined bycomparing their in vitro activity, and in vivo activity in animalmodels. Methods for the extrapolation of effective dosages in mice, andother animals, to humans are known to the art; for example, see U.S.Pat. No. 4,938,949. Useful dosages of Type IV PDE inhibitors are knownto the art. For example, see, U.S. Pat. No. 5,877,180, Col. 12.

[0183] Generally, the concentration of the compound(s) of formula (I) ina liquid composition, such as a lotion, will be from about 0.1-25% wt-%,preferably from about 0.5-10 wt-%. The concentration in a semi-solid orsolid composition such as a gel or a powder will be about 0.1-5 wt-%,preferably about 0.5-2.5 wt-%.

[0184] The amount of the compound, or an active salt or derivativethereof, required for use in treatment will vary not only with theparticular salt selected but also with the route of administration, thenature of the condition being treated and the age and condition of thepatient and will be ultimately at the discretion of the attendantphysician or clinician.

[0185] In general, however, a suitable dose will be in the range of fromabout 0.5 to about 100 μg/kg, e.g., from about 10 to about 75 μg/kg ofbody weight per day, such as 3 to about 50 μg per kilogram body weightof the recipient per day, preferably in the range of 6 to 90 μg/kg/day,most preferably in the range of 15 to 60 μg/kg/day.

[0186] The compound is conveniently administered in unit dosage form;for example, containing 5 to 1000 μg, conveniently 10 to 750 μg, mostconveniently, 50 to 500 μg of active ingredient per unit dosage form.

[0187] Ideally, the active ingredient should be administered to achievepeak plasma concentrations of the active compound of from about 0.1 toabout 10 nM, preferably, about 0.2 to 10 nM, most preferably, about 0.5to about 5 nM. This may be achieved, for example, by the intravenousinjection of a 0.05 to 5% solution of the active ingredient, optionallyin saline, or orally administered as a bolus containing about 1-100 μgof the active ingredient. Desirable blood levels may be maintained bycontinuous infusion to provide about 0.01-5.0 μg/kg/hr or byintermittent infusions containing about 0.4-15 μg/kg of the activeingredient(s).

[0188] The desired dose may conveniently be presented in a single doseor as divided doses administered at appropriate intervals, for example,as two, three, four or more sub-doses per day. The sub-dose itself maybe further divided, e.g., into a number of discrete loosely spacedadministrations; such as multiple inhalations from an insufflator or byapplication of a plurality of drops into the eye. For example, it isdesirable to administer the present compositions intravenously over anextended period of time following the insult that gives rise toinflammation.

[0189] The ability of a given compound of the invention to act as anA_(2A) adenosine receptor agonist (or antagonist) may be determinedusing pharmacological models which are well known to the art, or usingtests described below.

[0190] The invention will be further described by reference to thefollowing detailed examples, which are given for illustration of theinvention, and are not intended to be limiting thereof.

DESCRIPTION OF PREFERRED EMBODIMENTS

[0191] All melting points were determined with a Thomas Hoover capillarymelting point apparatus and are uncorrected. Nuclear magnetic resonancespectra for proton (¹H NMR) were recorded on a 300 MHz GEspectrophotometer. The chemical shift values are expressed in ppm (partsper million) relative to tetramethylsilane. For data reporting,s=singlet, d=doublet, t=triplet, q=quartet, and m=multiplet. Massspectra were measured on a Finnigan LcQ Classic. High resolution massspectrometry (HRMS) data was provided by the Nebraska Center for MassSpectrometry. Analytical HPLC was done on a Waters 2690 SeparationModule with a Waters Symmetry C8 (2.1×150 mm) column operated at roomtemperature. Compounds were eluted at 200 μL/min with 70:30acetonitrile:water, containing 0.5% acetic acid, with UV detection at214 nm using a Waters 486 Tunable Detector. Preparative HPLC wasperformed on a Shimadzu Discovery HPLC with a Shim-pack VP-ODS C₁₈(20×100 mm) column operated at room temperature. Compounds were elutedat 30 mL/min with a gradient 20-80% of water (containing 0.1% TFA) tomethanol over 15 minutes with UV detection at 214 nm using a SPD10A VPTunable detector. All final compounds presented here were determined tobe greater than 98% pure by HPLC. Flash chromatography was performed onSilicyle 60A gel (230-400 mesh) or using reusable chromatography columnsand system from RT Scientific, Manchester NH. Analytical thin-layerchromatography was done on Merck Kieselgel 60 F254 aluminum sheets.Preparative thin-layer chromatography was done using 1000 micronAnaltech Uniplate with silica gel. All reactions were done under anitrogen atmosphere in flame-dried glassware unless otherwise stated.

[0192] General method 1: Preparation of alkynyl cyclohexanols

[0193] To a solution of about 10 mmol of the appropriate cyclohexanonein about 50 mL of THF is added to about 60 mL (30 mmol) of 0.5 Methynylmagnesium bromide in THF. The solution is allowed to stir atabout 20° C. for about 20 hours. After the starting material had beenconsumed, monitored by TLC, the reaction is quenched with about 5 mL ofwater, filtered over a plug of sand and silica, washed with EtOAc, andevaporated to yield a yellow oil. Usually the oil contained two spots onTLC with 20% EtOAc/Hexanes, which are visualized with Vanillin. Usuallythese two products are the different isomers formed by theaxial/equatorial addition of the alkyne to the ketone. The compounds arepurified via flash chromatography using 10% EtOAc/Hexanes to provideclear oils or white solids in a yield of about 50-80 %.

[0194] General method 2: Preparation of propargylpiperadines/piperazines.

[0195] To a solution of of the appropriate piperazine/piperadine(about10.0 mmol), in about 20 mL acetonitrile, is added about 12.0 mmol ofpropargyl bromide (80% stabilized in toluene) and about 50.0 mmol ofanhydrous potassium carbonate. The reaction mixture is filtered, andevaporated to dryness. The resiude is taken up in about 50 mL ofdichloromethane/water and the organic layers removed. The aqueous layeris washed with an additional 3×25 mL dichloromethane. The organic layeris dried using anhydrous sodium sulfate, filtered, and concentrated toprovide the crude product, which is purified using columnchromatography.

[0196] General method 3: Preparation of modifiedpiperadines/piperazines.

[0197] To about 100 mg of the appropriate Boc-protectedpiperazine/piperadine is added 2-4 mL of neat TFA. The solution isallowed to stir for 6 hours. The TFA is removed under reduced pressureto yield a yellow oil. This oil is taken up in about 10 mL ofdichloromethane to which is added 10-fold excess of TEA and 3equivalents of the appropriate acyl chloride. The yellow solution isallowed to stir at room temperature for about 12 hours, after which timethe solvents are removed and the product purified using a 1.1×30 cm 14 gcolumn from Robert Thompson Scientific with a 5%-30% gradient of ethylacetate/hexanes.

[0198] General method 4: Preparation of 2-AAs (2-alkynyladenosines).

[0199] A flame-dried 25 mL round bottom under nitrogen is charged with5-(6-amino-2-iodo-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide (2-Iodoadenosine) (about 40 mg) (X═CH₃CH₂NHC(O)—) anddissolved in about 2 mL of DMF. The appropriate alkyne (approx. 0.1 mL)is then added followed by about 4 mL of acetonitrile and about 0.1 mL ofTEA. All three solvents had been degassed with nitrogen for at least 24hours. To this solution is added 5 mole percent Pd(PPh₃)₄ and 6 mole %copper iodide. The yellowish solution is allowed to stir for 24 hours atroom temperature, or until complete by HPLC. If the reaction is notcomplete at this time, additional catalyst, CuI, and TEA are added.After the reaction is complete, the solvents are removed underhigh-vacuum and the red/black residue taken back up in a small amount ofDMF. This solution is added to a preparative silica TLC plate (Analtech1000 microns, 20 cm×20 cm) and eluted first with 120 mL of 40%Hexanes/CH₂Cl₂, and then again after addition of 40 mL of MeOH. The UVactive band (usually yellow in color) in the middle of the plate iscollected, slowly washed with 4×25 mL 20% MeOH/CH₂Cl₂, and concentrated.This product is then purified by RP-HPLC.

[0200] Preparation 1:[(2R,3R,4R,5R)-3,4-diacetyloxy-5-(2-amino-6-oxohyropurin-9-yl)oxolan-2-yl]methylacetate (6.2).

[0201] A suspension of 113 g (0.4 mol) of dry guanosine (6.1), aceticanhydride (240 mL, 2.5 mol), dry pyridine (120 mL) and dry DMF (320 mL)was heated for 3.75 hours at 75° C. without allowing the temperature toexceed 80° C. The clear solution was then transferred to a 3L Erlenmyerflask and filled with 2-propanol. Upon cooling the solution to roomtemperature crystallization was initiated and allowed to proceed at 4°C. overnight. The white solid filtrate was filtered, washed with2-propanol and recrystallized from 2-propanol to provide 6.2 (96%). ¹HNMR (300 Mhz, CDCl₃) 8.20 (s, 1H, H-8), 6.17 (d, J=5.41 Hz, 1 H, H-1 )5.75 (t, J=5.39 Hz, 1H, H-2), 5.56 (t, J=5.0, H-3), 4.41 (m, 3H, H-4,5),2.14 (s, 3H, Ac), 2.11 (s, 3H, Ac), 2.10 (s, 3H, Ac). ¹³C NMR (300 MHz,CD₃OD) 171.0, 170.3, 1702, 157.7, 154.8, 152.4, 136.7, 117.7, 85.5,80.4, 73.0, 71.3, 64.0, 31.3, 21.2, 21.0.

[0202] Preparation 2:[(2R,3R,4R,5R)-3,4-diacetyloxy-5-(2-amino-6-chloropurin-9-yl)oxolan-2-yl]methylacetate (6.3).

[0203] To a 1 L flask was added 80 g (0.195 mol)[(2R,3R,4R,5R)-3-4-diacetyloxy-5-(2-amino-6-oxohyropurin-9-yl)oxolan-2-yl]methyl acetate (6.2), tetramethylammonium chloride (44 g, 0.4 mol),anhydrous acetonitrile (400 mL) and N,N-dimethlaniline (25 mL). Theflask was placed in an ice salt bath and cooled to 2° C. To thissolution was added dropwise POCl₃ (107 mL 1.15 mol) at a rate thatmaintained the temperature below 5° C. (45 minutes). The flask was thenremoved from the ice bath, outfitted with a condenser, placed in an oilbath and allowed to reflux for 10 minutes. The solution changed to ared/brown color. The solvent was removed under reduced pressure to yieldan oily residue which was transferred to a beaker containing 1000 g ofice and 400 mL of CHCl₃ and allowed to stir for 1.5 hours to decomposeany remaining POCl₃. The organic phase was removed and the aqueous phaseextracted with 3×50 mL of CHCl₃ and pooled with the organic phase. Thepooled organic layeres were back extracted with 50 mL of water followedby stirring with 200 mL of saturated NaHCO₃. The organic layer wasfurther extracted with NaHCO₃ until the aqueous extract was neutral(2×). The organic layer was finally extracted with brine and dried overMgSO₄ for 16 hours. To the solution was added 800 mL of 2-propanol afterwhich the solution was concentrated under reduced pressure. To the oilysolid was added 200 mL of 2-propanol and the solution was refrigeratedovernight. The crystalline product was filtered, washed, and allowed todry overnight to give 6.3 (77%). ¹H NMR (300 MHz, CD₃OD) 8.31 (s, 1H,H-8), 7.00 (s, 2H, NH₂) 6.06 (d, J=5.8 Hz, 1H, H-1), 5.83 (t, J=6.16 Hz,1H, H-2), 5.67 (m, 1H, H-3), 4.29 (m, 3H, H-4,5), 2.07 (s, 3H, Ac), 1.99(s, 3H, Ac), 1.98 (s, 3H, Ac). ¹³C NMR (300 MHz, CD₃OD) 171.0, 170.4,170.2, 160.8, 154.6, 150.8, 142.2, 124.5, 85.8, 80.6, 72.8, 71.2, 63.9,21.4, 21.3, 21.1.

[0204] Preparation 3:[(2R,3R,4R,5R)-3,4-diacetyloxy-5-(6-chloro-2-iodopurin-9-yl)oxolan-2-yl]methylacetate (6.4).

[0205] Isoamyl nitrite (5 mL, 37 mmol) was added to a mixture of 5.12 g(12 mmol)

[0206][(2R,3R,4R,5R)-3-,4-diacetyloxy-5-(2-amino-6-chloropurin-9-yl)oxolan-2-yl]methyl acetate (6.3), I₂ (3.04 g, 12 mmol), CH₂I₂ (10 mL, 124 mmol), and CuI(2.4 g, 12.6 mmol) in THF (60 mL). The mixture was heated under refluxfor 45 minutes and then allowed to cool to room temperature. To thissolution was added 100 ml of saturated Na₂S₂O₃. This step removed thereddish color. The aqueous layer was extracted 3× with chloroform, whichwas pooled, dried over MgSO₄, and concentrated under reduced pressure.The product was then purified over a silica gel column using CHCl₃-MeOH(98:2) to collect

[0207][(2R,3R,4R,5R)-3,4-diacetyloxy-5-(6-chloro-2-iodopurin-9-yl)oxolan-2-yl]methylacetate (6.4) (80% crystallized from EtOH). ¹H NMR (300 MHz, CDCl₃) 8.20(s, 1H H-8), 6.17 (d, J=5.41 Hz, 1H, H-1), 5.75 (t, J=5.39 Hz, 1H, H-2),5.56 (t, J=5.40Hz, 1H, H-3), 4.38 (m, 3H, H-4,5), 2.14 (s, 1H, Ac), 2.11(s, 1H, Ac), 2.10 (s, 1H, Ac).

[0208] Preparation 4:(4S,2R,3R,5R)-2-(6-amino-2-iodopurin-9-yl)-5-(hydroxy-methyl)oxolane-3,4-diol(6.5).

[0209] To a flask containing 6.0 g (11.1 mmol)

[0210][(2R,3R,4R,5R)-3,4-diacetyloxy-5-(6-chloro-2-iodopurin-9-yl)oxolan-2-yl]methylacetate (6.4) was added 100 ml of liquid NH₃ at −78° C. and the solutionwas allowed to stir for 6 hours. After which time it was allowed to cometo room temperature overnight with concurrent evaporation of the NH₃ toyield a brown oil. The product was crystallized from hot isopropanol toprovide 6.5 (80%), m.p. 143-145° C., r.f.=0.6 in 20% MeOH/CHCl₃. ¹H NMR(300 MHz, DMSO-d₆) 8.24 (s, 1H), 7.68 (s, 2H), 5.75 (d, J=6.16, 1H),5.42 (d, J=5.40 Hz, 1H), 5.16 (d, J=4.62 Hz, 1H), 4.99 (t, J=5.39 Hz,1H), 4.67 (d, J=4.81 Hz, 1H), 4.06 (d, J=3.37 Hz, 1H), 3.89 (m, 1H),3.54 (m, 2H).

[0211] Preparation 5:[(1R,2R,4R,5R)-4-(6-amino-2-iodopurin-9-yl)-7-7-dimethyl-3,6,8-trioxabicyclo[3.3.0]oct-2-yl]methan-1-ol(6.6).

[0212] To a solution of 2.0 g (5.08 mmol)

[0213](4S,2R,3R,5R)-2-(6-amino-2-iodopurin-9-yl)-5(hydroxymethyl)oxolane-3,4-diol(6.6) in 100 mL acetone was added 9.6 g of p-toluenesulfonic acid and 5ml of dimethoxypropane. The reaction was stirred at room temperature for1 hour. Solid NaHCO₃, 15 g, was added to the solution. The slurry wasstirred for an additional 3 hours. The residue was filtered and washed2× with EtOAc. The filtrate was then concentrated under reducedpressure. The residue was chromatographed on a silica gel column withMeOH-CHCl₃ (1:99) to give 6.6 (72%) as a solid, m.p. 185-187° C. ¹H NMR(300 MHz, DMSO-d₆) 8.22 (s, 1H, H-8), 7.69 (s, 2H), NH₂), 6.00 (d,J=2.70 Hz, 1H, H-1), 5.21 (m, 1H, H-2), 5.07 (bs, 1H, OH), 4.88 (m, 1H,H-3), 4.13 (m, 1H, H-4), 3.47 (m, 2H, H-5), 1.49 and 1.28 (s, 3H,C(CH₃)₂).

[0214] Preparation 6:(2S,1R,4R,5R)-4-(6-amino-2-iodopurin-9-yl)-7,7-dimethyl-3,6,8-trioxabicyclo[3.3.0]octane-2-carboxylicacid (6.7).

[0215] To a stirred solution of 1.6 g (3.7 mmol) of

[0216][(1R,2R,4R,5R)-4-(6-amino-2-iodopurin-9-yl)-7-7-dimethyl-3,6,8-trioxabicyclo[3.3.0]oct-2-yl]methan-1-ol (6.6) in 200 mL of H₂O was added 0.60 g of KOHand, dropwise, a solution of 1.70 g (10.8 mml) of KMnO₄ in 50 mL of H₂O.The mixture was placed in the dark at room temperature for 2-4 days. Thereaction mixture was then cooled to 5-10° C. and decolorized by asolution of 4 mL of 30% H₂O₂ in 16 mL of water, while the temperaturewas maintained below 10° C. using an ice-salt bath. The mixture wasfiltered through Celite and the filtrate was concentrated under reducedpressure to about 10 mL and then acidified to pH 4 with 2N HCl. Theresulting precipitate was filtered off and washed with ether to yield6.7 (70%) after drying as a white solid, m.p. 187-190 C. ¹H NMR (300MHz, DMSO-d₆) 8.11 (s, 1H, H-8), 7.62 (s, 2H, NH₂), 7.46 (s, 1H, COOH),6.22 (s, 1H, H-1), 5.42 (d, J=5.71 Hz, 1H, H-2), 5.34 (d, J=6.16 Hz, 1H,H-3), 4.63 (s, 1H, H-4), 1.46 and 1.30 (s, 3H, C(CH₃)₂).

[0217] Preparation 7:(2S,3S,4R,5R)-5-(6-amino-2-iodopurin-9-yl)-3,4-dihydroxyoxolane-2-carboxylicacid (6.8).

[0218] A solution of 1.72 g (3.85 mmol) of

[0219](2S,1R,4R,5R)-4-(6-amino-2-iodopurin-9-yl)-7,7-dimethyl-3,6,8-trioxabicyclo[3.3.0]octane-2-carboxylicacid (6.7) in 80 mL of 50% HCOOH was stirred at 80° C. for 1.5 hours.The reaction mixture was evaporated under reduced pressure, dissolved inH₂O, and the solvent was evaporated again. This process was repeateduntil there was no odor of formic acid in the residue. Recrystallizationfrom water provided 1.33 g (85%) 6.8 as a white solid, m.p. 221-223° C.,dec. ¹H NMR (300 MHz, DMSO-d₆) 8.31 (s, 1H, H-8), 7.68 (s, 2H, NH₂),5.90 (d, J=6.55 Hz, 1H, H-1H), 4.42(m, 1H, H-2), 4.35 (d, J=2.31 Hz, 1H,H-4), 4.22 (m, 1H, H-3).

[0220] Preparation 8:[(2S,3S,4R,5R)-5-(6-amino-2-iodopurin-9-yl)-3,4-dihydroxyoxolan-2-yl]-N-ethylcarboxamide(6.9).

[0221] To a cooled (5° C.) and stirred solution of 1.29 g (3.17 mmol) of

[0222](2S,3S,4R,5R)-5-(6-amino-2-iodopurin-9-yl)-3,4-dihydroxyoxolane-2-carboxylicacid (6.8) in 150 mL of absolute ethanol was added dropwise 1.15 mL ofice-cooled SOCl₂. The mixture was stirred at room temperature overnightand then brought to pH 8 with saturated aqueous NaHCO₃. The mixture wasfiltered, and then the filtrate was concentrated under reduced pressureto yield a white solid which was dried and then redissolved in 20 mL ofdry ethylamine at −20° C. for 3 hours and then at room temperatureovernight. The reaction mixture was diluted with absolute ethanol, andthe precipitated product was filtered off and washed with dry ether toprovide 530 mg (72%) of 6.9 as a pure solid, m.p. 232-234° C. ¹H NMR(300 MHz, DMSO-d₆) 8.34 (s, 1H, H-8), 8.12 (t, 1H, NH), 7.73 (s, 2H,NH₂), 5.85, (d, J=6.93 Hz, 1H, H-1), 4.54 (m, 1H, H-2), 4.25 (d, J=1.92Hz, 1H, H-4), 4.13 (m, 1H, H-3), 3.28 (m, 2H, CH₂CH₃), 1.00 (t, J=7.2Hz, 3H, CH₂CH₃).

[0223] Preparation 9:[4-(tert-Butyl-dimethyl-silanyloxymethyl)-cyclohexyl]-methanol (83).

[0224] To a 100 mL-flask containing 79 (4.0 g, 27.8 mmol) in DMF (40 mL)was added TBDMSCl (3.56 g, 23.6 mmol) and imidazole (3.79 g, 55.6 mmol).The reaction was allowed to stir at 25° C. for 16 hours after which timesaturated aqueous LiBr (50 mL) was added and the reaction extracted withether (2×50 mL). The ether layers were pooled and extracted again withLiBr (2×35 mL). The ether layer became clear. The ether layer was thenconcentrated in vacuo and the product purified by flash chromatography,on a silica gel column, eluting with 1:2 ether/petroleum ether to yield83 (3.80 g, 62%) as a homogenous oil. ¹H NMR (CDCl₃) δ3.46 (d, J=6.2 Hz,2 H), 3.39 (d, J=6.2 Hz, 2 H), 1.95-1.72 (m, 4 H), 1.65 (m, 1 H), 1.40(m, 1 H), 1.03-0.89 (m, 4 H), 0.88 (s, 9 H), 0.04 (s, 6 H; ¹³C NMR(CDCl₃) δ69.2, 69.1, 41.2, 41.1, 29.5, 26.5, 18.9, −4.8;. APCI m/z (relintensity) 259 (MH⁺, 100).

[0225] Preparation 10: Toluene-4-sulfonic acid4-(tert-butyl-dimethyl-silanyloxymethyl)-cyclohexylmethyl ester (84).

[0226] To a 100 mL-flask containing 83 (3.4 g, 13.2 mmol) in CHCl₃ (30mL) was added tosyl chloride (3.26 g, 17.1 mmol) and pyidine (3.2 mL,39.6 mmol). The reaction was allowed to stir at 25° C. for 14 hoursafter which time the reaction was concentrated in vacuo to yield a wetwhite solid. To this solid was added ether (50 mL) and the solid wasfiltered and subsequently washed with additional ether (2×50 mL). Theether layers were pooled, concentrated in vacuo to yield a clear oilwhich was purified by flash chromatography, on a silica gel column,eluting with 1:4 ether/petroleum ether to yield 84 (4.5 g, 83 %) as awhite solid. ¹H NMR (CDCl₃) δ7.78 (d, J=7.7, 2 H), 7.33 (d, J=7.7 Hz, 2H), 3,81 (d, J=6.2 Hz, 2H), 3.37 (d, J=6.2, 2 H), 2.44 (s, 3 H),1.95-1.72 (m, 4 H), 1.65 (m, 1 H), 1.40 (m, 1 H), 1.03-0.89 (m, 4 H),0.88 (s, 9 H), 0.04 (s, 6 H); ¹³C NMR (CDCl₃) δ145.1, 133.7, 130.3,128.4, 75.8, 68.9, 40.7, 38.0, 29.1, 26.5, 22.1, 18.9, −4.9; APCI m/z(rel intensity) 413 (MH⁺, 100).

[0227] Preparation 11: (4-Prop-2-ynyl-cyclohexyl)-methanol (86).

[0228] A 3-neck 250 mL-flask equipped with a gas inlet tube and dry-icecondenser was cooled to −78° C. and charged with liquid ammonia (40 mL).To the reaction mixture was added lithium wire (600 mg, 86.4 mmol)generating a deep blue solution. The mixture was allowed to stir for 1hour. Acetylene, passed through a charcoal drying tube, was added to theammonia until all the lithium had reacted and the solution turnedcolorless, at which time the flow of acetylene was stopped, theacetylene-inlet tube and condenser removed and the flask outfitted witha thermometer. DMSO (20 mL) was added and the ammonia evaporated with awarm water bath until the mixture reached a temperature of 30° C. Thesolution was stirred at this temperature for 2 hours until the solutionstopped bubbling. The mixture was cooled to 5° C. and compound 84 (11.25g, 27.3 mmol), in DMSO (10 mL), was added. The temperature wasmaintained at 5° C. The mixture was allowed to stir at 5° C. for 0.5hours. Then the solution was gradually warmed to room temperature andstirred for an additional 18 hours. The brown/black reaction mixture waspoured slowly over ice (300 g) and extracted with ether (4×100 mL),dried with anhydrous sodium sulfate, and concentrated in vacuo to yielda yellow oil. The oil was subsequently dissolved in THF (200 mL) andchanged to a brownish color upon addition of TBAF hydrate (11.20 g, 35.5mmol). The solution was allowed to stir for 24 hours under N₂atmosphere. After stirring, the reaction was quenched with water (200mL) and extracted with ether (3×100 mL). The ether extracts werecombined and concentrated in vacuo. The crude product was purified bychromatography, on a silica gel column, eluting with 1:1 ether/petroleumether to yield 86 (3.91 g, 93%) as a yellow oil. ¹H NMR (CDCl₃) δ3.45(d, J=6.2, 2 H), 2.10 (d, J=6.2, 2 H), 1.9 (s, 1 H), 1.94-1.69 (m, 4 H),1.52-1.34 (m, 2 H), 1.16 -0.83 (m, 4 H); ¹³C NMR (CDCl₃) δ83.8, 69.5,69.0, 40.8, 37.7, 32.3, 29.7, 26.5.

[0229] Preparation 12: (4-prop-2-ynylcyclohexyl)methyl acetate (87).

[0230] To a solution of 960 mg (6.31 mmol) of 86 in 6 mL DMF was added0.62 mL (7.57 mmol) pyridine and 0.78 mL (8.27mmol) acetic anhydride.The reaction was allowed to stir overnight at room temperature. After 16hours, starting material still remained. The reaction mixture was heatedat 75° C. for 3 hours. The solvent was removed under reduced pressure toyield a yellow oil which was purified by flash chromatography, on silicagel, eluting with 1:3 ether/petroleum ether to yield 1.12 g (91%) of 87as an oil. ¹H NMR (CDCl₃) δ3.87 (d, J=6.2Hz, 2 H), 2.06 (d, J=4.3 Hz, 2H), 2.03 (s, 3 H), 1.98-1.93 (m, 1 H), 1.92-1.83 (m, 2 H), 1.83-1.74 (m,2 H), 1.63-1.36 (m, 2 H), 1.12-0.90 (m, 4 H); ¹³C NMR (CDCl₃) δ171.7,83.7, 69.9, 69.6, 37.4, 37.3, 32.1, 29.7, 26.5, 21.4; APCI m/z (relintensity) 195 (M⁺, 30), 153 (M⁺, 70), 135 (M⁺, 100).

[0231] Preparation 13: 4-prop-2-ynyl-cyclohexanecarboxylic acid (88).

[0232] A solution of chromium trioxide (600 mg, 6.0 mmol) in 1.5 M H₂SO₄(2.6 mL, 150 mmol) was cooled to 5° C. and added to a solution of 86(280 mg, 1.84 mmol) in acetone (15 mL). The mixture was allowed to warmto room temperature and allowed to stir overnight. Isopropanol (4 mL)was added to the green/black solution, which turned light blue after 1hr. After adding water (15 mL), the solution was extracted with CHCl₃(6×25 mL). The organic layers were pooled and concentrated in vacuo toyield a white solid. The solid was dissolved in ether (50 mL) andextracted with 1 M NaOH (2×30 mL). The basic extracts were pooled,acidified w/10% HCl, and re-extracted with ether (3×30 mL). The etherlayers were combined, dried with sodium sulfate and concentrated invacuo to yield a white solid. The product was recrystallized fromacetone/water to yield 88 (222 mg, 73%) as white needles: mp 84-85° C.;¹H NMR (CDCl₃) δ2.30-2.23 (m, 1H), 2.17-2.11 (m, 2 H), 2.07-2.03 (m, 2H), 1.97-1.91 (m, 3H), 1.51-1.39 (m, 3 H), 1.13-1.01 (m, 2 H); ¹³C NMR(CDCl₃) δ182.5, 83.8, 69.6, 40.7, 37.7, 32.3, 29.6, 26.5; APCI m/z (relintensity) 165 (M⁻, 100).

[0233] Preparation 14: Methyl 4-prop-2-ynylcyclohexanecarboxylate (89).

[0234] To a solution of 88 (240 mg, 1.45 mmol) in 7:3 CH₂Cl₂:MeOH (10mL) was added TMS Diazomethane (2.0 M in hexanes) (0.9 mL, 1.8 mmol) in0.2 ml aliquots until the color remained yellow. The reaction wasallowed to stir for an additional 0.25 hours at room temperature. Afterstirring, glacial acetic acid was added dropwise until the solutionbecame colorless. The reaction was concentrated in vacuo to an oil whichwas purified by flash chromatography on silica gel using ether:petroleumether (1:9) to yield 89 (210 mg, 80%) as a clear oil. ¹H NMR (CDCl₃)δ3.60 (s, 3H), 2.25-2.13 (m, 1 H), 2.08-1.94 (m, 3 H), 1.95-1.90 (m, 2H), 1.49-1.31 (m, 3 H), 1.10-0.93 (m, 2 H); ¹³C NMR (CDCl₃) δ176.7,83.3, 69.8, 51.9, 43.4, 36.7, 31.9, 29.2, 26.3; APCI m/z (rel intensity)181 (MH⁺, 100).

[0235] Preparation 15: Trans[4-(1-Propargyl)cyclohexylmethyl] methylcarbonate (90).

[0236] Yield: 345 mg, 81%. ¹H NMR (CDCl₃) δ0.98-1.07, 1.40-1.52,1.57-1.70, 1.78-1.93 (4× m, 10H, cyclohexyl), 1.96 (t, 1H, acetylene),2.10 (dd, 2H, —C₆H₁₀CH₂CCH), 3.78 (s, 3H, —OCH₃), 3.96 (d, —C₆H₁₀CH₂O—).

[0237] Preparation 16: Trans[4-(1-Propargyl)cyclohexylmethyl] iso-butylcarbonate (91).

[0238] Yield: 433 mg, 83%. ¹H NMR (CDCl₃) δ0.95 (d, 4H, —OCH₂CH(CH₃)₂),0.98-1.09, 1.40-1.51, 1.57-1.70, 1.78-1.93 (4× m, 10H, cyclohexyl),1.94-2.04 (m, 1H, —OCH₂CH(CH₃)₂), 1.96 (t, 1H, acetylene), 2.10 (dd, 2H,—C₆H₁₀CH₂CCH), 3.91, 3.95 (2× d, 4H, —OCH₂CH(CH₃)₂, —C₆H₁₀CH₂O—).

[0239] Preparation 17: Trans[4-(1-Propargyl)cyclohexylmethyl] benzylcarbonate (92).

[0240] Yield: 340 mg, 69%. ¹H NMR (CDCl₃) δ0.97-1.08, 1.40-1.49,1.55-1.69, 1.77-1.93 (4× m, 10H, cyclohexyl), 1.96 (t, 1H, acetylene),2.10 (dd, 2H, —C₆H₁₀CH₂CCH), 3.98 (d, —C₆H₁₀CH₂O—), 5.15 (s, 2H,—OCH₂Ph), 7.33-7.40 (m, 5H, Ar).

[0241] Preparation 18:4-(Toluene-4-sulfonyloxymethyl)-piperidine-1-carboxylic acid tert-butylester (JR3215).

[0242] A solution of N-Boc-4-piperidinemethanol, 5.0 g (23.2 mmol) inchloroform, 50 mL, was prepared. Toluene sulfonyl chloride, 5.75 g (30.2mmol), in 5.6 mL of pyridine (69.6 mmol) was added. The solution wasstirred under nitrogen allowed to stir for 24 hours. Standard workup andchromatographic purification provided the title compound. Yield 6.0 g

[0243] Preparation 19: (R)-1-Ethynyl-(R)-3-methyl-cyclohexanol(JR3217A), (S)-1-Ethynyl-(R)-3-methyl-cyclohexanol (JR3217B).

[0244] To a solution of 1.0 g (8.9 mmol) (R)-(+)-3-methyl-cyclohexanonein 50 mL of THF was added 54 mL (26.7 mmol) of 0.5 M ethynylmagnesiumbromide in THF. The solution was allowed to stir at 20° C. for 20 hours.Analysis by TLC indicated that the starting material had been consumed.The reaction was quenched with 5 mL of water, filtered over a plug ofsand and silica, washed with EtOAc, and evaporated to yield 1.15 g of ayellow oil containing two spots (r.f.'s 0.33 (minor, JR3217A) and 0.25(major, JR3217B), 20% EtOAc/Hexanes) which were visualized withVanillin. The compound was purified via flash chromatography using 10%EtOAc/Hexanes (225 mL silica) to provide JR3217A and JR3217B.

[0245] Preparation 20: 1-Prop-2-ynyl-piperidine-2-carboxylic acid methylester (JR3249).

[0246] The title compound was prepared starting with 4.0 g (22.3 mmol)of methylpipecolinate hydrochloride according to general method 2.

[0247] Preparation 21: 1-Prop-2-ynyl-piperidine-4-carboxylic acid methylester (JR3245).

[0248] To a solution of methyl isonipecotate 3.5 g (24,4 mmol, 3.30 mL)in 100 mL dichloromethane was added TEA (1.5 eq, 36.6 mmol, 5.1 mL),propargyl bromide (3.0 eq, 73.2 mmol, 6.5 ml), at room temperature for36 hrs. The reaction was quenched with 35 mL water to yield to provide aclear solution. The solution was extracted with dichloromethane 2×25 mL,dried with Na2SO4, and the solvent evaporated to provide a yellow oil.r.f. (40% EtOAc/Hexanes) 0.26 stains faint white with Vanillin, startingmaterial r.f. 0.05 stains yellow with Vanillin. The product appearedpure after extraction.

[0249] Preparation 22: 1-Prop-2-ynyl-piperidine-4-carboxylic acid ethylester (JR3271).

[0250] The title compound was prepared starting with 2.0 g (12.7 mmol)of ethyl isonipecotate according to general method 2.

[0251] Preparation 23: 4-Prop-2-ynyl-piperazine-1-carboxylic acidtert-butyl ester (JR3275).

[0252] To a solution of 10.0 g (54.8 mmol) of tert-butyl-1-piperazinecarboxylate in 60 mL acetonitile was added 5.20 mL (60.4 mmol) propargylbromide and 37.9 g (274 mmol) anhydrous potassium carbonate. Additionalpropargy bromide, 1.5 mL, was added after stirring for 36 hours at roomtemperature. The residue was evaporated to dryness. Dichloromethane, 50mL, and water, 50 mL, were added. The reaction mixture was extractedwith CH₂Cl₂, 4×40 mL, dried over magnesium sulfate, and evaporate toprovide a brown oil. The oil was dissolved in dichloromethane and purifywith a RT Scientific system using hexane/ethyl acetate gradient to yield5.5 g (46%) of yellow oil, which ultimately crystallized upon standing.

[0253] Preparation 24: 4-Cyanomethyl-piperazine-1-carboxylic acid ethylester (JR3287).

[0254] To a solution of 3 g (19.0 mmol) of ethyl N-piperazinecarboxylatein 25 mL of CH₃CN was added 1.57 g (1.32 mL 20.1 mmol) of2-chloroacetonitrile and 15.6 g (95 mmol) K₂CO₃.1½H₂O. The suspensionwas stirred at room temperature for 16 hours. The reaction was analyzedusing TLC (35% Ethyl acetate/Hexanes, product r.f. 0.38 vs. sm r.f. of0.02). The analysis indicated the reaction was complete. The goldenyellow solution was evaporated to dryness. The residue was extractedwith CH₂C₁₂/H₂O, dried with MgSO₄, and concentrated.

[0255] Preparation 25:5-Prop-2-ynyl-2,5-diaza-bicyclo[2.2.1]heptane-2-carboxylic acidtert-butyl ester (JR4013).

[0256] The title compound was prepared starting with 500 mg (2.52 mmol)of 2,5-Diaza-bicyclo[2.2.1]heptane-2-carboxylic acid tert-butyl esteraccording to general method 2.

[0257] Preparation 26: 1-Cyclohexyl-4-prop-2-ynyl-piperazine (JR4019).

[0258] The title compound was prepared starting with 3 g (17.9 mmol) of1-cyclohexylpiperazine according to general method 2

[0259] Preparation 27: 1-Prop-2-ynyl-piperazine (JR4029).

[0260] To a flame-dried 25 mL round bottom flask under nitrogen wasadded 2.1 g of 4-Prop-2-ynyl-piperazine-1-carboxylic acid tert-butylester. To this solid was added 5 mL of 98% TFA in 1 mL portions. Thesolution turned wine red, bubbled and smoked. The additional portions ofTFA were added when this activity subsided. After the third portion ofTFA had been added only minimal bubbling occurred. The solution wasallowed to stir under nitrogen at room temperature for an additionalhour and evaporated under reduced pressure to yield the product as athick red syrup. Assumed quantitative yield of 1.16 g. The residue wassuspended in 20 mL dichloromethane and used immediately without furtherpurification for the preparation of compounds JR4031, JR4033, andJR4035.

[0261] Preparation 28: 4-Prop-2-ynyl-piperazine-1-carboxylic acid methylester (JR4031).

[0262] The title compound was prepared starting with 385 mg (3.1 mmol)of JR4029 and using methylchloroformate according to general method 3.

[0263] Preparation 29: 4-Prop-2-ynyl-piperazine-1-carboxylic acidisobutyl ester (JR4035).

[0264] The title compound was prepared starting with 385 mg (3.1 mmol)of JR4029 and using isobutylchloroformate according to general method 3.

[0265] Preparation 30:3,3-Dimethyl-1-(4-prop-2-ynyl-piperidin-1-yl)-butan-1-one (JR4041).

[0266] The title compound was prepared starting with tert-butyl ester(JR3257) and using tert-butylacetylchloride according to general method3.

[0267] Preparation 31: 1-(4-Prop-2-ynyl-piperazin-1-yl)-ethanone(JR4043).

[0268] The title compound was prepared starting with 385 mg (3.1 mmol)of JR4029 and using acetyl chloride according to general method 3.

[0269] Preparation 32: Piperidine-1,4-dicarboxylic acid mono-tert-butylester.

[0270] To a solution of piperidine-4-carboxylic acid (10 g, 77.5 mmol)and potassium carbonate (21.4 g, 155 mmol) in 150 mL of water wasprepared. A solution of di-tert-butyl dicarbonate (16.9 g, 77.5 mmol) in40 mL of THF was added dropwise via addition funnel at 0° C. Thereaction was allowed to warm to room temperature gradually over 30minutes and stirred for an additional 4 hours. The THF was removed underreduced pressure and the aqueous phase extracted with 50 mL of ether.The aqueous phase was then adjusted to pH 2 with 10% HCl and extractedwith EtOAc, 4×50 mL. The combined organic layers were dried overanhydrous sodium sulfate, filtered, and concentrated in vacuo to yield17.2 g (97%) of JR3183 as a white solid. Rf=0.2 (35% EtOAc/Hexanesstained w/vanillin). ¹H NMR (CDCl₃) δ11.83 (s, 1 H), 3.98 (d, J=11.8 Hz,2H), 2.83 (t, J=11.8, 2H), 2.46 (m, 1 H), 1.88 (d, J=12.9 hz, 2 H), 1.2(m, 2 H), 1.42 (s, 9 H). ¹³C NMR (CDCl₃) δ180.0, 154.8, 79.8, 42.9,40.8, 28.3, 27.7. APCI m/z (rel intensity) M⁻228.2 (100).

[0271] Preparation 33:

[0272] The following intermediate compounds are prepared using thegeneral method 1 described herein and the appropriate startingmaterials.

[0273] (R)-1-Ethynyl-3-tert-butyl-cyclohexanol (JR3255A),(S)-1-Ethynyl-3-tert-butyl-cyclohexanol (JR3255B).

[0274] Toluene-4-sulfonic acid 4-prop-2-ynyl-cyclohexylmethyl ester(JR3077).

[0275] 1-Ethyl-4-prop-2-ynyl-cyclohexane (JR3083).

[0276] 1-(4-Prop-2-ynyl-cyclohexyl)-ethanone (JR3115).

[0277] 1,1-Dicyclohexyl-prop-2-yn-1-ol (JR3127).

[0278] 1-Cyclohexyl-prop-2-yn-1-ol (JR3129).

[0279] 4-Ethyl-1-ethynyl-cyclohexanol (JR3143).

[0280] 1-Ethynyl-3-methyl-cyclohexanol.

[0281] 1-Ethynyl-3,3,5,5-tetramethyl-cyclohexanol (JR3151).

[0282] 1-Ethynyl-4-phenyl-cyclohexanol (JR3153).

[0283] 1-Ethynyl-2-methyl-cyclohexanol (JR3167B)

[0284] 4-tert-Butyl-1-ethynyl-cyclohexanol (JR3191).

[0285] 1-Ethynyl-3,3-dimethyl-cyclohexanol (JR3193).

[0286] Piperidine-1,4-dicarboxylic acid 1-tert-butyl ester 4-methylester (JR3195).

[0287] 4-Hydroxymethyl-piperidine-1-carboxylic acid tert-butyl ester(JR3199).

[0288] 4-Prop-2-ynyl-piperazine-1-carboxylic acid ethyl ester (JR3211).

[0289] 4-Prop-2-ynyl-piperidine-1-carboxylic acid tert-butyl ester(JR3257).

[0290] 4-Prop-2-ynyl-piperidine-1-carboxylic acid ethyl ester (JR3267B).

[0291] 2-(4-Prop-2-ynyl-piperazin-1-yl)-pyrimidine (JR3277).

[0292] 1-(4-Prop-2-ynyl-piperidin-1-yl)-ethanone (JR4037).

[0293] 2,2-Dimethyl-1-(4-prop-2-ynyl-piperidin-1-yl)-propan-1-one(JR4039).

EXAMPLE 14-{3-[6-Amino-9-(5-ethylcarbamoyl-3,4-dihydroxytetrahydro-furan-2-yl)-9H-purin-2-yl]-prop-2-ynyl}-cyclohexanecarboxylicacid (109)

[0294]

[0295] The reaction of 110 with five equivalents of LiOH in THF/waterfor 6 hours gave 109 (7 mg, 72%) as a white solid which was crystallizedfrom MeOH/H₂O(0.1% TFA) after purification by reverse phase HPLC. ¹H NMR(DMSO-d6) δ8.70 (s, 1 H), 8.41 (s, 1 H), 7.62 (s, 2 H), 5.89 (d, J=7.25Hz, 1 H), 4.53 (m, 1 H), 4.27 (s, 1 H), 4.08 (d, J=3.6 Hz, 1 H), 2.29(d, J=6.4 Hz, 2 H), 2.15-1.99 (m, 1 H), 1.92-1.76 (m, 4 H), 1.52-1.38(m, 1 H), 1.38-1.19 (m, 2 H), 1.02 (t, J=6.3 Hz 3 H); ¹³C NMR (DMSO-d6)176.7, 169.2, 155.6, 148.9, 145.2, 141.6, 119.0, 87.7, 85.0, 84.6, 81.6,73.1, 71.9, 43.2, 35.9, 33.3, 31.2, 28.3, 25.6, 15.0. HRMS (FAB) m/z474.2196 [(M+H)⁺ cacld for C₂₂H₂₉N₆O₆ 474.2182].

EXAMPLE 24-{3-[6-Amino-9-(5-ethylcarbamoyl-3,4-dihydroxytetrahydro-furan-2-yl)-9H-purin-2-yl]-prop-2-ynyl}-cyclohexanecarboxylic acid methylester (110)

[0296]

[0297] The reaction of 89 with 2-IodoNECA under the general conditionsdescribed above provided 110 (74 mg, 60%) as a white solid. ¹H NMR(CD₃OD) δ8.23 (s, 1 H), 5.92 (d, J=7.7 Hz, 1 H), 4.69-4.65 (dd, J=7.7Hz, 4.6 Hz, 1 H), 4.40 (s, 1 H), 4.24 (d, J=4.6 Hz, 1 H), 3.59 (s, 3 H),3.49-3.31 (m, 2 H), 2.31 (d, J=6.6Hz, 2 H), 2.10-2.09 (m, 1 H),2.01-1.89 (m, 4 H), 1.61-1.32 (m, 5 H), 1.13 (t, J=7.3 Hz, 3 H); ¹³C NMR(CD₃OD) δ177.1, 171.1, 156.3, 149.3, 146.7, 142.4, 119.7 89.6, 86.0,85.5, 81.6, 74.0, 72.2, 51.2, 43.2, 36.8, 34.2, 31.8, 28.9, 26.2, 14.4;HRMS (FAB) m/z 487.2325 [(M+H)⁺ cacld for C₂₃H₃₁N₆O₆ 487.2305].

EXAMPLE 3 Acetic acid 4-{3-[6-amino-9-(5-ethylcarbamoyl-3,4-dihydroxy-tetrahydrofuran-2-yl)-9H-purin-2-yl]-prop-2-ynyl}-cyclohexylmethylester (111)

[0298]

[0299] The reaction of 87 with 2-IodoNECA under the general conditionsdescribed above gave 111 (78 mg, 62%) as a white solid. ¹H NMR (CD₃OD)δ8.22 (s, 1 H), 5.92 (d, J=8.1 Hz, 1 H), 4.70-4.66 (dd, J=8.1 Hz, 4.6Hz, 1 H), 4.40 (d, J=1.2Hz, 1 H), 4.25-4.23 (dd, J=4.6 Hz, 1.2 Hz, 1 H),3.83 (d, J=6.5, 2 H), 3.53-3.31 (m, 2 H), 2.29 (d, J=6.5 Hz, 2 H), 1.97(s, 3 H), 1.93-1.89 (m, 2 H), 1.79-1.75 (m, 2 H), 1.64-1.42 (m, 2 H),1.12 (t, J=7.3 Hz, 3 H), 1.09-0.91 (m, 4 H); ¹³C NMR (CD₃OD) δ172.0,171.2, 156.2, 149.3, 146.7, 142.5, 119.7, 89.6, 86.3, 85.5, 81.5, 74.0,72.2, 69.6, 37.4, 37.2, 34.2, 32.1, 29.4, 26.4, 19.9, 14.5; HRMS (FAB)m/z 501.2469 [(M+H)⁺ cacld for C₂₄H₃₃N₆O₆ 501.2462].

EXAMPLE 45-{6-Amino-2-[3-(4-hydroxymethyl-cyclohexyl)-prop-1-ynyl]-purin-9-yl}-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide (112)

[0300]

[0301] The reaction of 86 (30 mg, 0.2 mmol) with 2-IodoNECA (28 mg, 0.07mmol) under the general conditions described above gave 112 (7 mg, 24%)as a white solid. ¹H NMR (CD₃OD) δ8.22 (s, 1 H), 5.92 (d, J=7.7 Hz, 1H), 4.70-4.66 (dd, J=7.7 Hz, 4.8 Hz, 1 H), 4.40 (d, J=1.2 Hz, 1 H),4.25-4.23 (dd, J=4.8 Hz, 1.2Hz, 1 H), 3.51-3.37 (m, 2 H), 3.31 (d,J=6Hz, 2 H), 2.30 (d, J=6.8 Hz, 2 H), 1.94-1.89 (m, 2 H), 1.83-1.78 (m,2 H), 1.64-1.42 (m, 2 H), 1.12 (t, J=7.3 Hz, 3 H), 1.09-0.91 (m, 4 H);¹³C NMR (CD₃OD) δ170.3, 155.4,148.5, 146.0, 141.6, 118.8, 88.7, 85.5,84.6, 80.6, 73.1, 71.3, 66.8, 39.6, 36.9, 33.3, 31.5, 28.6, 25.6, 13.5;HRMS (FAB) m/z 459.2373 [(M+H)⁺ cacld for C₂₂H₃₁N₆O₅ 459.2356].

EXAMPLE 55-{6-Amino-2-[3-(4-ethylcarbamoyl-cyclohexyl)-prop-1-ynyl]-purin-9-yl}-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide (JR3037)

[0302]

[0303] To a sealed tube containing 5 mL of freshly distilled ethylaminewas added 10 mg (0.02 mmol) of ATL146e. The flask was sealed and allowedto stir at 60° C. for 80 hours. After this time the reaction was onlyabout 50% complete by HPLC. The vessel was cooled to 0° C., opened, andthe ethylamine was removed in vacuo to yield 4.5 mg (73%) of JR3037 as awhite solid and the recovery of 4.0 mg of starting material after theresidue was purified by RP-HPLC. ¹H NMR (CD₃OD-d₄) δ. ¹³C NMR (CD₃OD-d₄)δ. APCI m/z (rel intensity) 500.8 (MH⁺, 100), 327.4(3).

EXAMPLE 65-{6-Amino-2-[3-(4-carbamoyl-cyclohexyl)-prop-1-ynyl]purin-9-yl}-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide (JR3055)

[0304]

[0305] To a sealed tube containing 10 mL of saturated MeOH/NH₃ solutionwas added 5 mg (0.01 mmol) of ATL146e. The flask was sealed and allowedto stir at 25° C. for 48 hours. The vessel was cooled to 0° C., opened,and the ammonia removed by bubbling N₂ for 1 hour. The remaining solventwas then removed in vacuo to yield 4.0 mg (83%) of JR3055 as a whitesolid after the residue was purified by RP-HPLC. ¹H NMR (CD₃OD-d₄) δ8.41(s, 1 H), 5.98 (d, J=7.2 Hz, 1H), 4.65 (dd, J=7.3 Hz, 4.8 Hz, 1 H), 4.41(d, J=2.0 Hz, 1 H), 4.28 (dd, J=4.6 Hz, 2.0 Hz, 1 H), 3.35 (m, 2 H),2,37 (d, J=6,4 Hz, 2 H) 2.10 (m, 1 H), 1.90 (m, _H), 1.53 (m, _H_), 1.23(m, ₁₃ H), 1,12 (t, J=7.3 Hz, 3 H). ¹³C NMR (CD₃OD-d₄) δ. APCI m/z (relintensity) 472.3 (MH⁺, 100), 299.4(10).

EXAMPLE 75-{6-Amino-2-[3-(4-methylcarbamoyl-cyclohexyl)-prop-1-ynyl]-purin-9-yl}-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide (JR3065)

[0306]

[0307] To a sealed tube containing 10 mL 2.0 M methylamine in methanolwas added 16.5 mg (0.03 mmol) of ATL146e. The flask was sealed andallowed to stir at 70° C. for 120 hours. The vessel was cooled to 0° C.,opened, and the solvent was removed in vacuo to yield 8.0 mg (48%) ofJR3065 as a white solid after the residue was purified by RP-HPLC. ¹HNMR (CD₃OD-d₄) δ. ¹³C NMR (CD₃OD-d₄) δ. APCI m/z (rel intensity) 486.3(MH⁺, 100), 313.4(35).

EXAMPLE 85-[6-Amino-2-(1-hydroxy-cyclopentylethynyl)-purin-9-yl]-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide (JR3135)

[0308]

[0309] The title compound was prepared using the appropriate startingmaterials and procedures described herein. The results are as follows:

[0310]¹H NMR (CD₃OD-d₄) δ8.48 (s, 1 H), 6.04 (d, J=6.9 Hz, 1 H), 4.72(dd, J=6.9 Hz, J=4.4 Hz, 1 H), 4.46 (d, J=2.3 Hz, 1 H), 4.33 (dd, J=4.6Hz, J=1.9 Hz, 1 H), 3.42 (m, 2 H), 2.04 (m, 4 H), 1.83, (m, 4 H), 1.16(t, J=7.3 Hz, 3 H). ¹³C NMR (CD₃OD-d₄) δ171.9, 155.3, 150.0, 144.3,120.6, 95.4, 90.6, 89.5, 86.2, 79.9, 74.9, 74.0, 70.5, 42.9, 35.3, 24.4,15.3. APCI m/z (rel intensity) 417.2 (MH⁺, 100), 399.4(85), 244.3(15),26.5(25). HRMS M⁺ actual 417.18864, observed 417.18880.

EXAMPLE 95-[6-Amino-2-(3,3-dicyclohexyl-3-hydroxy-prop-1-ynyl)-purin-9-yl]-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide (JR3139)

[0311]

[0312] The title compound was prepared using the appropriate startingmaterials and procedures described herein. The results are as follows:

[0313]¹H NMR (CD₃OD-d₄) δ8.57 (s, 1 H), 6.09 (d, J=6.6 Hz, 1 H), 4.77(dd, J=6.7, Hz, J=4.8 Hz, 1 H), 4.46 (d, J=2.3 Hz, 1 H), 4.37 (dd, J=4.6Hz, J=2.3 Hz, 1 H), 3.42 (m, 2 H) 1.80 (m, 13 H), 1.28 (m, 9 H), 1.13(t, J=7.3 Hz, 3 H). ¹³C NMR (CD₃OD-d₄) δ. APCI m/z (rel intensity) 527.3(MH⁺, 60), 509.5(100), 354.4(5), 336.5(5), 279.5(8). HRMS M⁺ actual527.29819, observed 527.29830

EXAMPLE 105-[6-Amino-2-(4-ethyl-1-hydroxy-cyclohexylethynyl)-purin-9-yl]-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide (JR3149)

[0314]

[0315] The title compound was prepared using the appropriate startingmaterials and procedures described herein. The results are as follows:

[0316]¹H NMR (CD₃OD-d₄) δ8.51 (s, 1 H), 6.06 (d, J=7.0 Hz, 1 H), 4.75(dd, J=6.4 Hz, J=4.9 Hz, 1 H), 4.46 (d, J=1.9 Hz, 1 H), 4.34 (dd, J=4.9Hz, J=2.1 Hz, 1 H), 3.42 (m, 2 H), 2.12 (d, J=11.9 Hz, 2 H), 1.80 (d,J=11.9 Hz, 2 H), 1.58 (t, J=12.1 Hz, 2 H), 1.28 (mn, 4 H), 1.15 (t,J=7.1 Hz, 3 H), 0.91 (t, J=7.1 Hz, 3 H). ¹³C NMR (CD₃OD-d₄) δ171.9,155.4, 150.0, 144.2, 143.8, 120.6, 94.5, 90.5, 86.1, 81.8, 74.9, 74.1,70.3, 40.5, 39.8, 35.3, 31.0, 30.2, 15.2, 12.0. APCI m/z (rel intensity)459.4 (MH⁺, 100), 441.4(60), 268.4(10). HRMS M⁺ actual 459.23559,observed 459.23550.

EXAMPLE 115-[6-Amino-2-(1-hydroxy-4-phenyl-cyclohexylethynyl)-purin-9-yl]-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide (JR3161)

[0317]

[0318] The title compound was prepared using the appropriate startingmaterials and procedures described herein. The results are as follows:

[0319]¹H NMR (CD₃OD-d₄) δ8.45 (s, 1 H), 7.26 (m, 4 H), 7.14 (m, 1 H),6.05 (d, J=7.3 Hz, 1 H), 4.80 (dd, J=7.3 Hz, J=4.8Hz, 1 H), 4.46 (d,J=1.6Hz, 1 H), 4.34 (dd, J=4.7 Hz, J=1.8 Hz, 1 H), 3.44 (m, 2 H), 2.58(m, 1 H), 2.23 (d, J=11.7 H, 2 H), 1.92 (m, 4 H), 1.78, (m, 2 H), 1.15(t, J=7.2 Hz, 3 H). ¹³C NMR (CD₃OD-d₄) δ. APCI m/z (rel intensity) 507.3(MH⁺, 100) 489.4(70), 334.3(5), 316.5(8). HRMS M⁺ actual 507.23559,observed 507.23580.

EXAMPLE 125-[6-Amino-2-(1-hydroxy-3,3,5,5-tetramethyl-cyclohexylethynyl)-purin-9-yl]-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide (JR3163)

[0320]

[0321] The title compound was prepared using the appropriate startingmaterials and procedures described herein. The results are as follows:

[0322]¹H NMR (CD₃OD-d₄) δ8.54 (s, 1 H), 6.04 (d, J=6.9 Hz, 1 H), 4.74(dd, J=6.9 Hz, J=5.0 Hz, 1 H), 4.46 (d, J=1.9 Hz, 1 H), 4.34 (dd, J=4.7Hz, J=1.9 Hz, 1 H), 3.44 (m, 2 H), 1.74 (s, 4 H), 1.13 (m, 17 H). APCIm/z (rel intensity) 487.3 (MH⁺, 75), 469.4(100), 296.4 (10).

EXAMPLE 135-[6-Amino-2-(1-hydroxy-2-methyl-cyclohexylethynyl)-purin-9-yl]-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide (JR3177A, JR3177B)

[0323]

[0324] The reaction of 1-Ethynyl-2-methyl-cyclohexanol (JR3169B) (100mg, 0.72 mmol) with 2-iodo-NECA (25 mg, 0.06 mmol) under the generalcoupling conditions gave JR3177A (8.0 mg) and JR3177B (8.2 mg) (overallyield 65%) as white solids after purification by a silica plug andRP-HPLC. JR3177A: ¹H NMR (CD₃OD-d₄) δ8.47 (s, 1 H), 6.05 (d, J=6.9 Hz, 1H), 4.77 (dd, J=6.9 Hz, J=4.9 Hz, 1 H), 4.45 (d, J=1.9 Hz, 1 H), 4.34(dd, J=4.6 Hz, J=2.1 Hz, 1 H), 3.41 (m, 2 H), 2.13 (d, J=12.7 Hz, 2 H),1.65 (m, 5 H), 1.32 (m, 2 H), 1.14 (t, J=7.0 Hz, 3 H), 1.13 (d, J=6.6Hz, 3 H). ¹³C NMR (CD₃OD-d₄) δ. APCI m/z (rel intensity) 445.3 (MH⁺,100), 427.4(80), 254.4(14). ¹H NMR (CD₃OD-d₄) δ8.49 (s, 1 H), 6.05 (d,J=6.9 Hz, 1 H), 4.78 (dd, J=6.4 Hz, J=4.9 Hz, 1 H), 4.45 (d, J=1.9 Hz, 1H), 4.34 (dd, J=4.6 Hz, J=1.6 Hz, 1 H), 3.42 (m, 2 H), 2.12 (d, J=12.3Hz, 2 H), 1.65 (m, 4 H), 1.35 (m, 4 H), 1.14 (t, J=7.3 Hz, 3 H), 1.12(d, J=6.6 Hz, 3 H). ¹³C NMR (CD₃OD-d₄) δ. APCI m/z (rel intensity) 445.7(MH⁺, 100), 427.3(35), 254.4(3.5).

EXAMPLE 145-[6-Amino-2-(1-hydroxy-3-methyl-cyclohexylethynyl)-purin-9-yl]-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide (JR3179)

[0325]

[0326] The reaction of 1-Ethynyl-3-methyl-cyclohexanol (JR3149B) (100mg, 0.72 mmol) with 2-iodo-NECA (25 mg, 0.06 mmol) under the generalcoupling conditions gave JR3179 (15.0 mg, 59%) as a white solid afterpurification by a silica plug and RP-HPLC. ¹H NMR (CD₃OD-d₄) δ8.49 (s, 1H), 6.06 (d, J=6.9 Hz, 1 H), 4.75 (dd, J=6.4 Hz, J=4.9 Hz, 1 H), 4.46(d, J=1.9 Hz, 1 H), 4.34 (dd, J=4.9 Hz, J=2.1 Hz, 1 H), 3.42 (m, 2 H),2.09 (d, J=12.3 Hz, 2 H), 1.73 (m, 4 H), 1.46 (m, 1 H), 1.23 (m, 1 H),1.169 (t, J=7.1 Hz, 3 H), 0.95 (d, J=6.2 Hz, 3 H), 0.89 (m, 1 H). ¹³CNMR (CD₃OD-d₄) δ. APCI m/z (rel intensity) 445.3 (MH⁺, 100), 427.4(40),254.4(4).

EXAMPLE 154-{3-[6-Amino-9-(5-ethylcarbamoyl-3,4-dihydroxytetrahydro-furan-2-yl)-9H-purin-2-yl]-prop-2-ynyl}-piperazine-1-carboxylicacid ethyl ester (JR3213)

[0327]

[0328] The title compound was prepared using the appropriate startingmaterials and procedures described herein. The results are as follows:

[0329]¹H NMR (CD₃OD-d₄) δ8.48 (s, 1 H), 6.00 (d, J=6.9 Hz, 1 H), 4.67(dd, J=6.5 Hz, J=5.0 Hz, 1 H), 4.42 (d, J=1.9 Hz, 1 H)), 4.39 (s, 2 H),4.35 (dd, J=4.7 Hz, J=1.9 Hz, 1 H), 4,13 (q,) 3.42 (m, 2 H), ¹³C NMR(CD₃OD-d₄) δ. APCI m/z (rel intensity) 503.4 (MH⁺, 100), 330.3 (6).

EXAMPLE 165-[6-Amino-2-(3-hydroxy-2-oxo-azepan-3-ylethynyl)purin-9-yl]-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide (JR3243A, JR3243B)

[0330]

[0331] 35 mg (0.081 mmol) IodoNECA (62 mg alkyne, 0.41 mmol), 2 ml DMF,4 ml Acetonitrile, 0.2 ml TEA, d(PPH3)4, CuI. Stirred overnight at roomtemperature (Nov. 29, 2001). R×n is tan w/ brown precipitate. TLC (20%MeOH/CH2Cl2) indicates r×n complete (r.f INECA=0.67, r.f product=0.45).Filtered mixture through celite, washed with 3×2 mL DMF, and evaporatedunder vacuum to brown oil. (solid precipitates out upon the addition ofMeOH, thus used DMF to load on prep plate).

[0332] The following compounds can be prepared by following the generalmethod 4 described herein and the appropriate intermediate compoundsdescribed herein.

EXAMPLE 17 N-Ethyl2-{3-[trans-4-(methoxycarbonyloxamethyl)-cyclohexyl]-1-propyn-1-yl}adenosine-5′-uronamide(ATL214)

[0333]

[0334] Yield 3.4 mg, 10%. ¹H NMR (CD₃OD) δ1.18 (t, 3H, —NHCH₂CH₃),1.03-1.20, 1.51-1.70, 1.79-1.85, 1.94-2.01 (4×m, 10H, cyclohexyl), 2.35(d, 2H, —C₆H₁₀CH₂CC—), 3.46 (m, 2H, —NHCH₂CH₃), 3.73 (s, 3H, —OCH₃),3.94 (d, 2H, —C₆H₁₀CH₂O—), 4.29 (dd, 1H, 3′-H), 4.45 (d, 1H, 4′-H), 4.72(dd, 1H, 2′-H), 5.97 (d, 1H, 1′-H), 8.27 (s, 1H, 8-H). APCI m/z 517.4(M+H⁺).

EXAMPLE 18 N-Ethyl2-{3-[trans-4-(isobutoxyoxycarbonyloxamethyl)-cyclohexyl]-1-propyn-1-yl}adenosine-5′-uronamide(ATL215)

[0335]

[0336] Yield 8.5 mg, 30%. ¹H NMR (CD₃OD) δ0.94 (d, 4H, —OCH₂CH(CH₃)₂),1.18 (t, 3H, —NHCH₂CH₃), 1.04-1.24, 1.54-1.72, 1.79-2.03 (3× m, 11H,cyclohexyl, —OCH₂CH(CH₃)₂), 2.38 (d, 2H, —C₆H₁₀CH₂CC—), 3.43 (m, 2H,—NHCH₂CH₃), 3.89, 3.94 (2× d, 4H, —C₆H₁₀CH₂O—, —OCH₂CH(CH₃)₂), 4.30 (dd,1H, 3′-H), 4.46 (d, 1H, 4′-H), 4.71 (dd, 1H, 2′-H), 6.00 (d, 1H, 1′-H),8.37 (br s, 1H, 8-H). APCI m/z 559.5 (M+H⁺).

EXAMPLE 19 N-Ethyl2-{3-[trans-4-(benzoxycarbonyloxamethyl)-cyclohexyl]-1-propyn-1-yl}adenosine-5′-uronamide(ATL216)

[0337]

[0338] Yield 1.0 mg, 3%. ¹H NMR (CD₃OD) δ1.17 (t, 3H, —NHCH₂CH₃),1.03-1.23, 1.52-1.71, 1.78-1.86, 1.93-2.02 (4× m, 10H, cyclohexyl), 2.35(d, 2H, —C₆H₁₀CH₂CC—), 3.45 (m, 2H, —NHCH₂CH₃), 3.97 (d, 2H,—C₆H₁₀CH₂O—), 4.29 (dd, 1H, 3′-H), 4.45 (d, 1H, 4′-H), 4.72 (dd, 1H,2′-H), 5.13 (s, 2H, —OCH₂Ph), 5.97 (d, 1H, 1′-H), 7.33-7.37(m, 5H, Ar),8.30 (br s, 1H, 8-H). APCI m/z 593.3 (M+H⁺).

EXAMPLE 204-{3-[6-Amino-9-(5-ethylcarbamoyl-3,4-dihydroxy-tetrahydro-furan-2-yl)-9H-purin-2-yl]-prop-2-ynyl}-cyclohexanecarboxylicacid 2-tert-butoxycarbonylamino-ethyl ester

[0339]

EXAMPLE 215-{6-Amino-2-[3-(4-dimethylaminomethyl-cyclohexyl)-prop-1-ynyl]-purin-9-yl}-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide (JR2023)

[0340]

EXAMPLE 224-{3-[6-Amino-9-(5-ethylcarbamoyl-3,4-dihydroxy-tetrahydro-furan-2-yl)-9H-purin-2-yl]-prop-2-ynyl}-cyclohexanecarboxylicacid 2-amino-ethyl ester (JR3033)

[0341]

EXAMPLE 234-{3-[6-Amino-9-(5-ethylcarbamoyl-3,4-dihydroxytetrahydro-furan-2-yl)-9H-purin-2-yl]-prop-2-ynyl}-1-methyl-cyclohexanecarboxylicacid methyl ester (JR3067A)

[0342]

EXAMPLE 244-{3-[6-Amino-9-(5-ethylcarbamoyl-3,4-dihydroxytetrahydro-furan-2-yl)-9H-purin-2-yl]-prop-2-ynyl}-1-methyl-cyclohexanecarboxylicacid methyl ester (JR3067B)

[0343]

EXAMPLE 255-{6-Amino-2-[3-(4-ethyl-cyclohexyl)-prop-1-ynyl]-purin-9-yl}-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide (JR3087)

[0344]

EXAMPLE 265-{2-[3-(4-Acetyl-cyclohexyl)-prop-1-ynyl]-6-aminopurin-9-yl}-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide (JR3119)

[0345]

EXAMPLE 275-(6-Amino-2-{3-[4-(1-hydroxy-ethyl)-cyclohexyl]-prop-1-ynyl}-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide

[0346]

EXAMPLE 285-[6-Amino-2-(1-hydroxy-2-methyl-cyclohexylethynyl)-purin-9-yl]-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide (JR3181A, JR3181B)

[0347]

EXAMPLE 295-[6-Amino-2-(1-hydroxy-3,3-dimethylcyclohexylethynyl)-purin-9-yl]-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide (JR3201B)

[0348]

EXAMPLE 305-[6-Amino-2-(4-tert-butyl-1-hydroxycyclohexylethynyl)purin-9-yl]-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide (JR3203)

[0349]

EXAMPLE 315-[6-Amino-2-(1-hydroxy-3-methylcyclohexylethynyl)purin-9-yl]-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide (JR3221)

[0350]

EXAMPLE 325-[6-Amino-2-(1-hydroxy-3-methylcyclohexylethynyl)purin-9-yl]-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide (JR3223)

[0351]

EXAMPLE 335-[6-Amino-2-(2-tert-butyl-1-hydroxycyclohexylethynyl)purin-9-yl]-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide (JR3227)

[0352]

EXAMPLE 341-{3-[6-Amino-9-(5-ethylcarbamoyl-3,4-dihydroxytetrahydro-furan-2-yl)-9H-purin-2-yl]-prop-2-ynyl}-piperidine-4-carboxylicacid methyl ester (JR3251)

[0353]

EXAMPLE 351-{3-[6-Amino-9-(5-ethylcarbamoyl-3,4-dihydroxytetrahydro-furan-2-yl)-9H-purin-2-yl]-prop-2-ynyl}-piperidine-2-carboxylicacid methyl ester (JR3253)

[0354]

EXAMPLE 364-{3-[6-Amino-9-(5-ethylcarbamoyl-3,4-dihydroxytetrahydro-furan-2-yl)-9H-purin-2-yl]-prop-2-ynyl}-piperidine-1-carboxylicacid tert-butyl ester (JR3259)

[0355]

EXAMPLE 374-{3-[6-Amino-9-(5-ethylcarbamoyl-3,4-dihydroxytetrahydro-furan-2-yl)-9H-purin-2-yl]-prop-2-ynyl}-piperidine-1-carboxylicacid ethyl ester (JR3269)

[0356]

EXAMPLE 381-{3-[6-Amino-9-(5-ethylcarbamoyl-3,4-dihydroxytetrahydro-furan-2-yl)-9H-purin-2-yl]-prop-2-ynyl}-piperidine-4-carboxylicacid ethyl ester (JR3279)

[0357]

EXAMPLE 394-{3-[6-Amino-9-(5-ethylcarbamoyl-3,4-dihydroxytetrahydro-furan-2-yl)-9H-purin-2-yl]-prop-2-ynyl}-piperazine-1-carboxylicacid tert-butyl ester (JR3281)

[0358]

EXAMPLE 405-{6-Amino-2-[3-(4-pyrimidin-2-yl-piperazin-1-yl)-prop-1-ynyl]-purin-9-yl}-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide (JR3283)

[0359]

EXAMPLE 415-[6-Amino-2-(3-piperazin-1-yl-prop-1-ynyl)purin-9-yl]-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide (JR3289)

[0360]

EXAMPLE 421-{3-[6-Amino-9-(5-ethylcarbamoyl-3,4-dihydroxytetrahydro-furan-2-yl)-9H-purin-2-yl]-prop-2-ynyl}-piperidine-4-carboxylicacid (JR3291)

[0361]

EXAMPLE 434-{3-[6-Amino-9-(5-ethylearbamoyl-3,4-dihydroxytetrahydro-furan-2-yl)-9H-purin-2-yl]-prop-2-ynyl}-piperidine-1-carboxylicacid methyl ester (JR4007)

[0362]

EXAMPLE 444-{3-[6-Amino-9-(5-ethylearbamoyl-3,4-dihydroxytetrahydro-furan-2-yl)-9H-purin-2-yl]-prop-2-ynyl}-piperidine-1-carboxylicacid isopropyl ester (JR4009)

[0363]

EXAMPLE 454-{3-[6-Amino-9-(5-ethylcarbamoyl-3,4-dihydroxytetrahydro-furan-2-yl)-9H-purin-2-yl]-prop-2-ynyl}-piperidine-1-carboxylicacid isobutyl ester (JR4011)

[0364]

EXAMPLE 465-{3-[6-Amino-9-(5-ethylcarbamoyl-3,4-dihydroxytetrahydro-furan-2-yl)-9H-purin-2-yl]prop-2-ynyl}-2,5-diazabicyclo[2.2.1]heptane-2-carboxylicacid tert-butyl ester (JR4015)

[0365]

EXAMPLE 475-(6-Amino-2-{3-[1-(3,3-dimethyl-butyryl)-piperidin-4-yl]-prop-1-ynyl}purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide (JR4047)

[0366]

EXAMPLE 485-(6-Amino-2-{3-[1-(2,2-dimethyl-propionyl)-piperidin-4-yl]prop-1-ynyl}-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide (JR4051)

[0367]

EXAMPLE 494-{3-[6-Amino-9-(5-ethylcarbamoyl-3,4-dihydroxytetrahydro-furan-2-yl)-9H-purin-2-yl]-prop-2-ynyl}-piperazine-1-carboxylicacid isobutyl ester (JR4049)

[0368]

EXAMPLE 505-{2-[3-(4-Acetyl-piperazin-1-yl)-prop-1-ynyl]-6-aminopurin-9-yl}-3,4-dihydroxytetrahydrofuran-2-carboxylicacid ethylamide (JR4053)

[0369]

EXAMPLE 51

[0370] The following compounds can be prepared by following the generalmethods described herein and the appropriate intermediate compounds:

EXAMPLE 52 Cell Culture and Membrane Preparation

[0371] Sf9 cells were cultured in Grace's medium supplemented with 10%fetal bovine serum, 2.5 μg/ml amphotericin B and 50 μg/ml gentamycin inan atmosphere of 50% N₂/50% O₂. Viral infection was performed at adensity of 2.5×10⁶ cells/mL with a multiplicity of infection of two foreach virus used. Infected cells were harvested 3 days post-infection andwashed twice in insect PBS (PBS pH 6.3). Cells were then resuspended inlysis buffer (20 mM HEPES pH 7.5, 150 mM NaCl, 3 mM MgCl₂, 1 mMβ-mercaptoethanol (BME), 5 μg/mL leupeptin, 5 μg/mL pepstatin A, 1 μg/mLaprotinin, and 0.1 mM PMSF) and snap frozen for storage at −80° C. Cellswere thawed on ice, brought to 30 mL total volume in lysis buffer, andburst by N₂ cavitation (600 psi for 20 minutes). A low-speedcentrifugation was performed to remove any unlysed cells (1000× g for 10minutes), followed by a high-speed centrifugation (17,000× g for 30minutes). The pellet from the final centrifugation was homogenized inbuffer containing 20 mM HEPES pH 8, 100 mM NaCl, 1% glycerol, 2 μg/mLleupeptin, 2 μg/mL pepstatin A, 2 μg/mL Aprotinin, 0.1 mM PMSF, and 10μM GDP using a small glass homogenizer followed by passage through a 26gauge needle. Membranes were aliquoted, snap frozen in liquid N₂, andstored at −80° C. Membranes from cells stably expressing the human A₁ AR(CHO K1 cells) or A₃ AR (HEK 293 cells) were prepared as described(Robeva et al., 1996).

EXAMPLE 53 Radioligand Binding Assays

[0372] Radioligand binding to recombinant human A_(A2) receptors in Sf9cell membranes was performed using either the radiolabeled agonist,¹²⁵I-APE (Luthin et al., 1995) or the radiolabeled antagonist,¹²⁵I-ZM241385 (¹²⁵I-ZM). To detect the high affinity, GTPγS-sensitivestate of A₁ and A₃ AR, we used the agonist, ¹²⁵I-ABA (Linden et al.,1985; Linden et al., 1993). Binding experiments were performed intriplicate with 5 μg (A_(2A)) or 25 μg (A₁ and A₃) membrane protein in atotal volume of 0.1 mL HE buffer (20 mM HEPES and 1 mM EDTA) with 1 U/mLadenosine deaminase and 5 mM MgCl₂ with or without 50 μM GTPγS.Membranes were incubated with radioligands at room temperature for threehours (for agonists) or two hours (for antagonists) in MilliporeMultiscreen® 96-well GF/C filter plates and assays were terminated byrapid filtration on a cell harvester (Brandel, Gaithersburg, Md.)followed by 4×150 μl washes over 30 seconds with ice cold 10 mMTris-HCl, pH 7.4, 10 mM MgCl₂. Nonspecific binding was measured in thepresence of 50 μM NECA. Competition binding assays were performed asdescribed (Robeva et al., 1996) using 0.5-1 nM ¹²⁵I-APE, ¹²⁵I-ZM241385,or ¹²⁵I-ABA. We found that it was sometimes important to change pipettetips following each serial dilution to prevent transfer on tips ofpotent hydrophobic compounds. The K_(i) values for competing compoundbinding to a single site were derived from IC₅₀ values with correctionfor radioligand and competing compound depletion as described previously(Linden, 1982).

[0373] Linden J (1982) Calculating the Dissociation Constant of anUnlabeled Compound From the Concentration Required to DisplaceRadiolabel Binding by 50%. J Cycl Nucl Res 8: 163-172.

[0374] Linden J, Patel A and Sadek S (1985) [¹²⁵I]Aminobenzyladenosine,a New Radioligand With Improved Specific Binding to Adenosine Receptorsin Heart. Circ Res 56: 279-284.

[0375] Linden J, Taylor H E, Robeva A S, Tucker A L, Stehle J H, RivkeesS A, Fink J S and Reppert S M (1993) Molecular Cloning and FunctionalExpression of a Sheep A₃ Adenosine Receptor With Widespread TissueDistribution. Mol Pharmacol 44: 524-532.

[0376] Luthin D R, Olsson R A, Thompson R D, Sawmiller D R and Linden J(1995) Characterization of Two Affinity States of Adenosine A_(A2)Receptors With a New Radioligand,2-[2-(4-Amino-3-[¹²⁵I]Iodophenyl)Ethylamino]Adenosine. Mol Pharmacol 47:307-313.

[0377] Robeva A S, Woodard R, Luthin D R, Taylor H E and Linden J (1996)Double Tagging Recombinant A₁- and A₂A-Adenosine Receptors WithHexahistidine and the FLAG Epitope. Development of an Efficient GenericProtein Purification Procedure. Biochem Pharmacol 51: 545-555.

[0378] Chemiluminescence Methods: Luminol enhanced chemiluminescence, ameasure of neutrophil oxidative activity, is dependent upon bothsuperoxide production and mobilization of the granule enzymemyeloperoxidase. The light is emitted from unstable high-energy oxygenspecies such as hypochlorous acid and singlet oxygen generated byactivated neutrophils.

[0379] Purified human neutrophils (2×106/ml) suspended in Hanks balancedsalt solution containing 0.1 % human serum albumin (HA), adenosinedeaminase (1U/mL) and rolipram (100 nM) were incubated (37 C.) in awater bath for 15 min with or without rhTNF(10U/ml). Followingincubation 100 L aliquots of the PMN were transferred to wells (Whitewalled clear bottom 96 well tissue culture plates Costar #3670; 2wells/condition) containing 501 HA and luminol (final concentration100M) with or without adenosine agonist (final agonist concentrations0.01-1000 nM). The plate was incubated 5 min (37 C.) and then fMLP (50 1in HA; final concentration 1M) was added to all wells.

[0380] Peak chemiluminescence was determined with a Victor 1420Multilabel Counter in the chemiluminescence mode using the WallacWorkstation software. Data are presented as peak chemiluminescence aspercent of activity in the absence of an adenosine agonist. The EC50 wasdetermined using PRISM software. All compounds were tested with PMNsfrom three separate donors. The results are summarized in Table 8. TABLE8 Binding Affinity And Selectivity For A_(2A) Agonists PMN PMN oxid +oxid roli A1/ A3/ (-log (-log Agonist A2A ± SEM A1 ± SEM A3 ± SEM A2AA2A EC50) EC50) ATL-146a 29.6 ± 1.2 189 ± 19 29 ± 10 6.4 1.0 6.04 7.72ATL-146e 0.5 ± 0.04 77.0 ± 12 45.0 ± 15 154.0 90.0 8.45 9.33 ATL-193 1.1± 0.2 71.0 ± 14 231.0 ± 91 64.5 210.0 8.51 9.46 ATL-2037 1.5 ± 0.01 19.0± 1 76.0 ± 6 12.6 50.3 7.49 8.7 NECA 2.0 ± 0.4 2.0 ± 1.4 32.0 ± 9 1.016.0 7.82 8.95 CCPA 11.0 ± 1.9 0.3 ± 0.1 65.0 ± 6 0.0 5.9 5.37 7.26CGS-21680 4.9 ± 0.3 316.0 ± 59 82.0 ± 18 64.5 16.7 7.52 8.55 CI-IBMECA18.3 ± 3.2 33.0 ± 9 2.4 ± 0.3 1.8 0.1 4.34 7.49 CPA 19.8 ± 3.2 0.4 ± 0.193.0 ± 7 0.02 4.7 4.03 7.06 IBMECA 6.3 ± 1.3 9.0 ± 0.8 1.5 ± 0.3 1.4 0.26.49 8.01 JR-2145 0.7 ± 0.16 28.3 ± 7.3 101.0 ± 22 42.2 150.7 7.19 8.57JR-2147 0.9 ± 0.38 17.5 ± 5.2 89.0 ± 21 19.0 96.7 7.13 8.57 JR-2171 1.2± 0.09 132.0 ± 13 62.0 ± 12 110.0 51.7 8.1 9.27 JR-3021 1.6 ± 0.5 107.0± 10 41.0 ± 1 66.9 25.6 7.58 8.58 JR-3023 3.8 ± 2 14.0 ± 0.8 61.0 ± 63.7 16.3 7.37 8.66 JR-3027 0.8 ± 0.1 23.4 ± 3.7 26.0 ± 9 28.2 31.3 8.469.46 JR-3031 13.8 ± 2.8 734.0 ± 126 126.0 ± 20 53.2 9.1 6.26 7.75JR-3033 1.6 ± 0.02 15.1 ± 3.9 26.0 ± 14 9.7 16.7 7.31 8.67 JR-3037 4.1 ±0.9 113.0 ± 11 93.0 ± 21 27.6 22.7 7.34 8.68 JR-3041 12.0 ± 4.8 7.1 ±2.2 82.0 ± 8 0.6 6.8 4.19 6.95 JR-3055 1.8 ± 0.2 36.0 ± 2 28.0 ± 3 20.015.6 7.5 8.64 JR-3063 0.3 ± 0.02 0.2 ± 0.03 1.3 ± 0.4 0.9 5.2 9.63 10.33JR-3065 3.6 ± 0.5 55.0 ± 13 28.0 ± 13 15.3 7.8 6.99 8.29 JR-3067A 1.3 ±0.8 99.0 ± 27 39.0 ± 3 76.2 30.0 7.65 8.7 JR-3067B 4.0 ± 0.2 121.0 ± 44212.0 ± 95 30.3 53.0 6.83 8.2 JR-3079 1.1 ± 0.33 0.5 ± 0.04 15.6 ± 3 0.414.1 7.85 9.11 JR-3081 0.9 ± 0.18 0.5 ± 0.09 14.7 ± 7.5 0.6 15.6 8.349.57 JR-3085 0.5 ± 0.18 1.0 ± 0.26 2.0 ± 1.3 2.0 4.1 8.41 9.6 JR-30872.2 ± 0.2 20.3 ± 4.5 17.8 ± 5.5 9.2 8.1 7.33 8.51 JR-3089 0.7 ± 0.05 7.1± 1.1 4.4 ± 11 10.8 6.7 9.14 9.9 JR-3101 0.9 ± 0.16 2.7 ± 0.23 6.2 ± 0.63.1 7.0 8.41 9.5 JR-3103 1.7 ± 0.21 2.0 ± 0.61 5.0 ± 2.3 1.2 3.0 7.959.03 JR-3119 2.4 ± 0.2 28.7 ± 3.3 18.6 ± 4.4 11.9 7.7 7.54 8.7 JR-31215.7 ± 0.6 52.3 ± 7.6 71.0 ± 16 9.3 12.6 6.98 8.31 JR-3135 0.8 ± 0.3 5.0± 1.3 3.5 ± 1.2 6.3 4.4 8.9 9.6 JR-3137 1.2 ± 0.35 13.5 ± 3.2 7.4 11.36.2 7.59 8.72 JR-3139 166.0 ± 42 801.0 ± 99 83.7 ± 30 4.8 0.5 5.39 6.75JR-3141 0.8 ± 0.28 9.7 ± 2.3 70.0 11.5 83.3 8.19 9.4 JR-3149 0.5 ± 0.053.8 ± 0.4 7.3 ± 1.9 7.0 13.5 8.66 9.63 JR-3159A 0.6 ± 0.06 0.9 ± 0.053.9 ± 1.5 1.4 6.6 8.43 9.34 JR-3159B 0.4 ± 0.07 2.0 ± 0.21 2.6 ± 0.3 5.77.4 9.44 10.36 JR-3161 0.9 ± 0.04 10.9 ± 0.5 113.0 ± 6 12.2 127.0 8.359.29 JR-3163 11.1 ± 2.8 784.0 ± 94 152.0 ± 62 70.6 13.7 6.27 7.67JR-3177A 1.2 ± 0.35 8.7 ± 0.07 6.7 ± 0.3 7.1 5.5 7.53 9.03 JR-3177B 0.7± 0.07 15.5 ± 0.7 7.5 ± 4.5 23.1 11.2 8.5 9.45 JR-3179 0.4 ± 0.05 8.5 ±0.3 12.6 ± 3.6 19.3 28.6 9.18 10.07 JR-3181A 1.2 ± 0.15 44.9 ± 17.2 69.7± 43 38.1 59.1 7.73 8.77 JR-3181B 0.7 ± 0.07 24.2 ± 6.2 38.9 ± 4.8 36.158.1 8.57 9.67 JR-3201B 1.6 ± 0.28 157.0 ± 14 23.6 ± 0.35 98.1 14.8 8.069.2 JR-3203 1.5 ± 0.22 14.7 ± 2.2 43.3 ± 18 9.5 28.1 7.85 9.11 JR-32053.4 ± 0.3 14.2 ± 1.08 31.6 ± 26.5 4.2 9.3 7.22 8.08 JR-3213 1.9 ± 0.415.8 ± 0.48 42.3 ± 18.3 8.3 22.3 8.46 9.11 JR-3221 5.08 ± 0.81 19.5 ±0.12 9.0 ± 5.5 3.8 1.8 7.3 8.21 JR-3223 0.92 ± 0.2 4.8 ± 0.3 0.7 ± 0.025.2 0.8 9.47 10.15 JR-3229 20.8 ± 2.5 234 15.3 ± 5.42 11.3 0.7 6.65 7.58JR-3227 25.1 ± 1.3 536 50 21.4 2.0 5.13 7.14 JR-3243A 11.9 ± 1.3 29.6 ±4.8 31.1 ± 7.2 2.5 2.6 6.42 7.65 JR-3243B 2.25 ± 0.35 12 ± 5 3.34 ± 0.485.3 1.5 7.81 8.91 JR-3251 6.02 ± 0.57 47.3 ± 24.2 58.5 ± 8 7.9 9.7 7.528.5 JR-3253 3.29 ± 0.49 13.8 ± 0.5 31.3 ± 9.3 4.2 9.5 7.36 8.42 JR-32590.9 ± 0.10 205 ± 2.6 45.6 ± 10 227.8 50.7 8.46 9.26 JR-3261 5.7 ± 1.847.3 ± 11.6 86.7 ± 3.75 8.3 15.2 6.4 7.6 JR-3269 1.26 ± 0.21 134 ± 12.568.6 106.0 54.4 8.65 JR-3279 3.82 ± 0.95 14.4 3.8 0.0 7.86 JR-3281 2.36± 0.50 9.15 ± 1.35 38 3.9 16.1 8.19 JR-3283 2.00 ± 0.25 15.3 ± 0.4 66.24.6 19.0 8.24 JR-4007 0.97 ± 0.33 34.6 ± 1 .4 65.2 35.7 67.2 8.77JR-4009 1.43 ± 0.23 167 ± 13.5 86 116.4 60.1 8.45 JR-4011 0.61 ± 0.0671.7 0.0 117.5 8.46 JR-4015 4.25 ± 1.0 0.0 0.0 ATL214 0.8 100 100 125125 2 .6 ATL215 4.8 20 2.8 ATL216 2.7 13 1.8 AB-1 0.71 ± 0.09 76.7 ± 8.94.9 ± 0.23 108.0 6.9 8.94 9.54 AB-3 2.55 ± 0.20 75.7 ± 9.2 7.8 ± 2.829.7 3.0 8.39 9.4 AB-5 5.48 ± 1.1 55.4 ± 4.8 12.5 ± 5.7 10.1 2.3 7.88.83 AB-6 5.8 ± 1.6 25.9 ± 3.5 11.1 ± 1.72 4.5 1.9 7.53 8.51 AB-8 1.20 ±0.05 36.9 ± 6.1 11.9 ± 1.86 30.8 9.9 7.76 8.72

EXAMPLE 54 Effect of A_(A2) Agonists on Neutrophil Oxidative Activity

[0381] A. Materials.

[0382] f-met-leu-phe (fMLP), luminol, superoxide dismutase, cytochromeC, fibrinogen, adenosine deaminase, and trypan blue were obtained fromSigma Chemical. Ficoll-hypaque was purchased from ICN (Aurora, Ohio),and Cardinal Scientific (Santa Fe, N. Mex.) and Accurate Chemicals andScientific (Westerbury, N.Y.). endotoxin (lipopolysaccharide; E. coliK235) was from List Biologicals (Campbell, Calif.). Hanks balanced saltsolution (HBSS), and limulus amebocyte lysate assay kit were fromBioWittaker (Walkersville, Md.). Human serum albumin (HSA) was fromCutter Biological (Elkhart, Ind.). Recombinant human tumor necrosisfactor-alpha was supplied by Dianippon Pharmaceutical Co. Ltd. (Osaka,Japan). ZM241385(4-(2-[7-amino-2-(2-furyl)[1,2,4]triazolo[2,3-a][1,3,5]triazin-5-ylamino]ethyl)phenol) was a gift from Simon Poucher, ZenecaPharmaceuticals, Cheshire, UK. Stock solutions (1 mM and 10 mM in DMSO)were made and stored at −20° C.

[0383] B. Human Neutrophil Preparation

[0384] Purified neutrophils (˜98% neutrophils and >95% viable by trypanblue exclusion) containing <1 platelet per 5 neutrophils and <50 pg/mlendotoxin (limulus amebocyte lysate assay) were obtained from normalheparinized (10 U/ml) venous blood by a one step Ficoll-hypaqueseparation procedure (A. Ferrante et al., J. Immunol. Meth., 36, 109(1980)).

[0385] C. Release of Inflammatory Reactive Oxygen Species from Primedand Stimulated Human Neutrophils Chemiluminescence

[0386] Luminol-enhanced chemiluminescence, a measure of neutrophiloxidative activity, is dependent upon both superoxide production andmobilization of the lysosomal granule enzyme myeloperoxidase. The lightis emitted from unstable high-energy oxygen species generated byactivated neutrophils. Purified neutrophils (5-10×10⁵/ml) were incubatedin Hanks balanced salt solution containing 0.1% human serum albumin (1ml) with the tested A_(A2) agonist with or without rolipram and with orwithout tumor necrosis factor-alpha (1 U/ml) for 30 minutes at 37° C. ina shaking water bath. Then luminol (1×10⁻⁴ M) enhanced f-met-leu-phe (1mcM) stimulated chemiluminescence was read with a Chronolog® Photometer(Crono-log Corp., Havertown, Pa.) at 37° C. for 2-4 minutes.Chemiluminescence is reported as relative peak light emitted (=height ofthe curve) compared to samples with tumor necrosis factor-alpha andwithout agonist or rolipram.

EXAMPLE 55 In vivo Rat Blood Pressure Experiments

[0387] Sprague-Dawley rats (mean weights, 250-300 grams) wereanthesthetized and jugular and carotid catheters are implantedipsilaterally and the animals are allowed to recover 24-48 hours. Priorto each experiment a baseline blood pressure reading is established for30 minutes with each drug injection being preceeded by a vehiclecontrol. Drugs are injected bolus I.V. through a jugular catheter in a200 microliter volume of saline and the catheter is flushed with anadditional 300 microliters of saline. To measure blood pressure, acentral line from the carotid catheter is attached to the pressuretransducer of a Digi-Med Blood Pressure Analyzer. Systolic pressure,diastolic pressure, mean pressure, and heart rate are all recorded inreal time at 30-60 second intervals. Data is recorded until mean bloodpressure has returned to baseline and remained constant for 20 minutes.The data is presented as a fraction of the mean blood pressure averagedover the 10 minutes immediately prior to drug injection. The bloodpressures are recorded and plotted over time as a means of determiningpotency of the compounds as well as biological half-life.

[0388] The results are illustrated in FIGS. 1-6.

EXAMPLE 56 In vivo Coronary Dog Flow Experiments

[0389] Fasted, adult mongrel dogs (mean weight, 24.8±0.6 kg; range, 20.9to 28.2 kg) were anaesthetized with sodium pentobarbital (30 mg·kg⁻¹),tracheally intubated, and mechanically ventilated with room air on arespirator (model 613, Harvard Apparatus) with positive end-expiratorypressure of 5 cm H₂O. The surgical preparation and instrumentation ofthe animals has been thoroughly described previously (Glover D. K. etal., Circulation 1996, 94, pages 1726-1732). Throughout each experiment,heart rate, mean arterial and left atrial pressures, ultrasonicallymeasured LCx flow, and dP/dt were continuously monitored and recorded ona 16-channel thermal array chart recorder (K2-G, Astro-med, Inc) anddigitised and stored on an IBM-compatible personal computer. Allexperiments were performed with the approval of the University ofVirginia Animal Care and Use Committee and were in compliance with theposition of the American Heart Association on the use of researchanimals. The compounds tested were intravenously administered by bolusinjection and the parameters above were measured and recorded.

[0390] The results are illustrated in FIGS. 7-16.

EXAMPLE 57 Liver I/R Injury Protocol

[0391] Mice were anesthetized by intraperitoneal injection of ketamine100 mg/kg and xylazine 10 mg/kg. Glycopyrrolate (Robinul-V) 0.05 mg/kgwas delivered subcutaneously before the operation. The ambienttemperature was controlled in the range of 24° C. to 26° C. Mice wereplaced on a 37° C. heat pad with their core temperature monitored by aTH-8 Thermalert Monitoring Thermometer (Physitemp) and maintained at36-37° C. by a TCAT-1A Temperature Control and Alarm Unit (Physitemp)during the entire procedure. After midline laparotomy, a microaneurysmclip was applied to the hepatic triad above the bifurcation to clamp theflow of the hepatic artery, portal vein, and bile duct. The peritoneumwas closed after superfusion of 200 μl of warm saline. After 60 minutesof ischemia, the peritoneum was reopened and the microaneurysm clip wasremoved. Immediately after reperfusion was initiated, each mousereceived a loading dose of ATL-146e (1 ug/kg) or vehicle in 200 uL warmsaline, and a primed Alzet osmotic minipump was placedintraperitoneally. The surgical wound was closed with metal staples.Mice were maintained on the heat pad to monitor and maintain bodytemperature until the anesthetic wore off.

[0392] Drug Administration.

[0393] Alzet osmotic minipumps (model 1003D; Alza Corp., Palo Alto,Calif., USA) were primed according to the manufacture's instruction inorder to release compounds shortly after implantation. A solutioncontaining ATL146e was prepared in non-nal saline and placed in osmoticminipumps to deliver 10 ng/kg/min. Minipumps containing vehicle orATL146e were implanted during operation.

EXAMPLE 58 Serum Enzyme Determination

[0394] Serum GPT (ALT) levels were measured using a Transaminase kit(505, Sigma). Briefly, 20 μL serum sample was mixed with 100 (Lpre-heated Alanine-α-KG substrate and incubated in a 37° C. water bathfor 30 minutes. Then we added 100 (L Sigma Color Reagent to the reactionand left it at room temperature for 20 minutes. We stopped the reactionwith 1.0 ml 0.4N sodium hydroxide solution. Absorbance of each sample at505 nm was measured and converted into SF unit/ml.

EXAMPLE 59 Tissue Myeloperoxidase Measurement

[0395] Mouse livers were removed after 24 hours reperfusion. The tissuewas immediately submerged in 10 volumes of ice-cold 50 mM KPO4 buffer,pH 7.4 and homogenized with a Tekmar tissue grinder. The homogenate wascentrifuged at 15,000× g for 15 minutes at 4° C., and the supernatantwas discarded. The pellet was washed twice, resuspended in 10 volumes ofice-cold 50 mM KPO4 buffer pH 7.4 with 0.5% hexadecyltrimethylammoniumbromide and then sonicated. The suspension was subjected to threefreeze/thaw cycles. Samples were sonicated for 10 seconds, andcentrifuged at 15,000× g for 15 minutes at 4° C. The supernatant wasadded to an equal volume of a solution consisting of o-dianisidine (10mg/ml), 0.3% H2O2, and 50 mM KPO4, pH 6.0. Absorbance was measured at460 nm over a period of five minutes.

[0396]FIG. 17 illustrates the longer duration of action of JR3223 vs. acontrol compound and ATL146e for liver tissue protection after anischemia/reperfusion injury. The test compounds were administered 6hours prior to I/R injury. Tissue protection is measured by amount ofSerum GPT present in the in a serum sample 24 hours later, with smallerGPT concentrations indicating better liver function.

[0397] All publications, patents, and patent documents are incorporatedby reference herein, as though individually incorporated by reference.The invention has been described with reference to various specific andpreferred embodiments and techniques. However, it should be understoodthat many variations and modifications may be made while remainingwithin the spirit and scope of the invention.

1. A compound having formula (I):

wherein Z is CR³R⁴R⁵ or NR⁴R⁵; each R¹ is independently hydrogen, halo,—OR^(a), —SR^(a), (C₁-C₈)alkyl, cyano, nitro, trifluoromethyl,trifluoromethoxy, C₃₋₈cycloalkyl, heterocycle,hetrocycle(C₁-C₈)alkylene-, aryl, aryl(C₁-C₈)alkylene-, heteroaryl,heteroaryl(C₁-C₈)alkylene-, —CO₂R^(a), R^(a)C(═O)O—, R^(a)C(═O)—,—OCO₂R^(a), R^(a)R^(b)NC(═O)O—, R^(b)OC(═O)N(R^(a))—, R^(a)R^(b)N—,R^(a)R^(b)NC(═O)—, R^(a)C(═O)N(R^(b))—, R^(a)R^(b)NC(═O)N(R^(b))—,R^(a)R^(b)NC(═S)N(R^(b))—, —OPO₃R^(a), R^(a)OC(═S)—, R^(a)C(═S)—,—SSR^(a), R^(a)S(═O)—, R^(a)S(═O)₂—, —N═NR^(a), or —OPO₂R^(a); each R²is independently hydrogen, halo, (C₁-C₈)alkyl, (C₃-C₈)cycloalkyl,heterocycle, heterocycle(C₁-C₈)alkylene-, aryl, aryl(C₁-C₈)alkylene-,heteroaryl, or heteroaryl(C₁-C₈)alkylene-; or R¹ and R² and the atom towhich they are attached is C═O, C═S or C═NR^(c): R⁴ and R⁵ together withthe atoms to which they are attached form a saturated or partiallyunsaturated, mono-, bicyclic- or aromatic ring having 3, 4, 5, 6, 7, 8,9 or 10 ring atoms optionally comprising 1, 2, 3, or 4 heteroatomsselected from non-peroxide oxy (—O—), thio (—S—), sulfinyl (—SO—),sulfonyl (—S(O)₂—) or amine (—NR^(a)—) in the ring; wherein any ringcomprising R⁴ and R⁵ is substituted with from 1 to 14 R⁶ groups; whereineach R⁶ is independently halo, —OR^(a), —SR^(a), (C₁-C₈)alkyl, cyano,nitro, trifluoromethyl, trifluoromethoxy, (C₁-C₈)cycloalkyl,(C₆-C₁₂)bicycloalkyl, heterocycle or hetrocycle (C₁-C₈)alkylene-, aryl,aryl (C₁-C₈)alkylene-, heteroaryl, heteroaryl(C₁-C₈)alkylene-,—CO₂R^(a), R^(a)C(═O)O—, R^(a)C(═O)—, —OCO₂R^(a), R^(a)R^(b)NC(═O)O—,R^(b)OC(═O)N(R^(a))—, R^(a)R^(b)N—, R^(a)R^(b)NC(═O)—,R^(a)C(═O)N(R^(b)), R^(a)R^(b)NC(═O)N(R^(b))—,R^(a)R^(b)NC(═S)N(R^(b))—, —OPO₃R^(a), R^(a)OC(═S)—, R^(a)C(═S)—,—SSR^(a), R^(a)S(═O)—, —NNR^(a),—OPO₂R^(a), or two R⁶ groups and theatom to which they are attached is C═O, C═S or; two R⁶ groups togetherwith the atom or atoms to which they are attached can form a carbocyclicor heterocyclic ring; R³ is hydrogen, halo, —OR^(a), —SR^(a),(C₁-C₈)alkyl, cyano, nitro, trifluoromethyl, trifluoromethoxy,(C₃-C₈)cycloalkyl, heterocycle, hetrocycle(C₁-C₈)alkylene-, aryl,aryl(C₁-C₈)alkylene-, heteroaryl, heteroaryl(C₁-C₈)alkylene-, —CO₂R^(a),R^(a)C(═O)O—, R^(a)C(═O)—, —OCO₂R^(a), R^(a)R^(b)NC(═O)O—,R^(b)OC(═O)N(R^(a))—, R^(a)R^(b)N—, R^(a)R^(b)NC(═O)—,R^(a)C(═O)N(R^(b))—, R^(a)R^(b)NC(═O)N(R^(b))—,R^(a)R^(b)NC(═S)N(R^(b))—, —OPO₃R^(a), R^(a)OC(═S)—, R^(a)C(═S)—,—SSR^(a), R^(a)S(═O)—, R^(a)S(═O)₂—, —NNR^(a), —OPO₂R^(a); or if thering formed from CR⁴R⁵ is aryl or hetreroaryl or partially unsaturatedthen R³ can be absent; each R⁷ is independently hydrogen, (C₁-C₈)alkyl,(C₃-C₈)cycloalkyl, aryl or aryl(C₁-C₈)alkylene, heteroaryl,heteroaryl(C₁-C₈)alkylene-; X is —CH₂OR^(a), —CO₂R^(a), —OC(O)R^(a),—CH₂OC(O)R^(a), —C(O)NR^(a)R^(b), —CH₂SR^(a), —C(S)OR^(a), —OC(S)R^(a),—CH₂OC(S)R^(a) or C(S)NR^(a)R^(b) or —CH₂N(R^(a))(R^(b)); wherein any ofthe alkyl, cycloalkyl, heterocycle, aryl, or heteroaryl, groups of R¹,R², R³, R⁶ and R⁷ is optionally substituted on carbon with one or more(e.g. 1, 2, 3, or 4) substituents selected from the group consisting ofhalo, —OR^(a), —SR^(a), (C₁-C₈)alkyl, cyano, nitro, trifluoromethyl,trifluoromethoxy, (C₃-C₈)cycloalkyl, (C₆-C₁₂)bicycloalkyl, heterocycleor hetrocycle(C₁-C₈)alkylene-, aryl, aryloxy, aryl (C₁-C₈)alkylene-,heteroaryl, heteroaryl(C₁-C₈)alkylene-, —CO₂R^(a), R^(a)C(═O)O—,R^(a)C(═O)—, —OCO₂R^(a), R^(a)R^(b)NC(═O)O—, R^(b)OC(═O)N(R^(a))—,R^(a)R^(b)N—, R^(a)R^(b)NC(═O)—, R^(a)C(═O)N(R^(b))—,R^(a)R^(b)NC(═O)N(R^(b))—, R^(a)R^(b)NC(═S)N(R^(b))—, —OPO₃R^(a),R^(a)OC(═S)—, R^(a)C(═S)—, —SSR^(a), R^(a)S(═O)_(p)—,R^(a)R^(b)NS(O)_(p)—, N═NR^(a), and —OPO₂R^(a); wherein any(C₁-C₈)alkyl, (C₃-C₈)cycloalkyl, (C₆-C₁₂)bicycloalkyl, (C₁-C₈)alkoxy,(C₁-C₈)alkanoyl, (C₁-C₈)alkylene, or heterocycle, is optionallypartially unsaturated; R^(a) and R^(b) are each independently hydrogen,(C₁-C₈)alkyl, or (C₁-C₈)alkyl substituted with 1-3 (C₁-C₈)alkoxy,(C₃-C₈)cycloalkyl, (C₁-C₈)alkylthio, amino acid, aryl,aryl(C₁-C₈)alkylene, heteroaryl, or heteroaryl(C₁-C₈)alkylene; or R^(a)and R^(b), together with the nitrogen to which they are attached, form apyrrolidino, piperidino, morpholino, or thiomorpholino ring; and R^(c)is hydrogen or (C₁-C₆)alkyl; m is 0 to about 8 and p is 0 to 2; providedthat when CR⁴R⁵ is a carbocyclic ring then at least one of R¹, R², or R³is a group other than hydrogen or at least one R⁶ group is a group otherthan —CH₂OH, —CO₂R^(a), R^(a)C(═O)O—, —R^(a)C(═O)OCH₂— orR^(a)R^(b)NC(═O)—; provided that m is at least 1 when Z is NR⁴R⁵; or apharmaceutically acceptable salt thereof.
 2. The compound of claim 1,wherein R¹ is hydrogen, —OH, —CH₂OH, —OMe, —OAc, —NH₂, —NHMe, —NMe₂ or—NHAc.
 3. The compound of claim 2, wherein R¹ is hydrogen, —OH, —OMe,—OAc, —NH₂, —NHMe, —NMe₂ or —NHAc.
 4. The compound of claim 3, whereinR¹ is hydrogen, OH, OMe, or NH₂.
 5. The compound of claim 4, wherein R¹is hydrogen, OH, or NH₂.
 6. The compound of claim 5, wherein R¹ ishydrogen or OH.
 7. The compound of claim 1, wherein R² is hydrogen,(C₁-C₈)alkyl, cyclopropyl, cyclohexyl or benzyl.
 8. The compound ofclaim 7, wherein R² is hydrogen, methyl, ethyl or propyl.
 9. Thecompound of claim 8, wherein R² is hydrogen or methyl.
 10. The compoundof claim 9, wherein R² is hydrogen.
 11. The compound of claim 1, whereinR¹, R² and the carbon atom to which they are attached is carbonyl (C═O).12. The compound of claim 1, wherein R³ is hydrogen, OH, OMe, OAc, NH₂,NHMe, NMe₂ or NHAc.
 13. The compound of claim 12, wherein R³ ishydrogen, OH, OMe, or NH₂.
 14. The compound of claim 13, wherein R³ ishydrogen, OH, or NH₂.
 15. The compound of claim 14, wherein R³ ishydrogen or OH.
 16. The compound of claim 1, wherein the ring comprisingR⁴, R⁵ and the atom to which they are connected is cyclopentane,cyclohexane, piperidine, dihydro-pyridine, tetrahydro-pyridine,pyridine, piperazine, decaline, tetrahydro-pyrazine, dihydro-pyrazine,pyrazine, dihydro-pyrimidine, tetrahydro-pyrimidine,hexahydro-pyrimidine, pyrazine, imidazole, dihydro-imidazole,imidazolidine, pyrazole, dihydro-pyrazole, and pyrazolidine.
 17. Thecompound of claim 16, wherein the ring comprising R⁴, R⁵ and the atom towhich they are connected is cyclopentane, cyclohexane, piperidine,dihydro-pyridine, tetrahydro-pyridine, pyridine, piperazine,tetrahydro-pyrazine, dihydro-pyrazine, pyrazine, dihydro-pyrimidine,tetrahydro-pyrimidine, hexahydro-pyrimidine, pyrazine, imidazole,dihydro-imidazole, imidazolidine, pyrazole, dihydro-pyrazole, andpyrazolidine.
 18. The compound of claim 17, wherein the ring comprisingR⁴ and R⁵ and the atom to which they are connected is, cyclohexane,piperidine or piperazine.
 19. The compound of claim 1, wherein R⁶ is(C₁-C₈)alkyl, or substituted (C₁-C₈)alkyl, —OR^(a), —CO₂R^(a),R^(a)C(═O)—, R^(a)C(═O)O—, R^(a)R^(b)N—, R^(a)R^(b)NC(═O)—, or aryl. 20.The compound of claim 19, wherein R⁶ is (C₁-C₈)alkyl, —OR^(a),—CO₂R^(a), R^(a)C(═O)—, R^(a)C(═O)O—, R^(a)R^(b)N—, R^(a)R^(b)NC(═O)—,or aryl.
 21. The compound of claim 20, wherein R⁶ is methyl, ethyl,butyl, OH, OR^(a), —CO₂R^(a), R^(a)C(O)—, OC(═O)CH₂CH₃, —CONR^(a)R^(b),NR^(a)R^(b) or phenyl.
 22. The compound of claim 21, wherein R⁶ is OH,OMe, methyl, ethyl, t-butyl, —CO₂R^(a), —CONR^(a)R^(b), OAc, NH₂, NHMe,NMe₂, NHEt or N(Et)₂.
 23. The compound of claim 22, wherein R⁶ ismethyl, ethyl, t-butyl, phenyl, —CO₂R^(a)—CONR^(a)R^(b), or —(═O)CR^(a).24. The compound of claim 23, wherein R⁶ is methyl, ethyl, —CO₂R^(a)—CONR^(a)R^(b), or OAc.
 25. The compound of claim 24, wherein R⁶ is—(CH₂)₁₋₂OR^(a), —(CH₂)₁₋₂C(═O)OR^(a), —(CH₂)₁₋₂OC(═O)R^(a),—(CH₂)₁₋₂C(═O)R^(a), —(CH₂)₁₋₂OCO₂R^(a), —(CH₂)₁₋₂NHR^(a),—(CH₂)₁₋₂NR^(a)R^(b), —(CH₂)₁₋₂OC(═O)NHR^(a), or—(CH₂)₁₋₂OC(═O)NR^(a)R^(b).
 26. The compound of claim 25, wherein R⁶ is—CH₂OH, —CH₂OAc, —CH₂OCH₃, —CH₂C(═O)OCH₃, —CH₂OC(═O)CH₃, —CH₂C(═O)CH₃,—CH₂OCO₂CH₃, —CH₂NH(CH₃), or —(CH₂)₁₋₂N(CH₃)₂.
 27. The compound of claim27, wherein R⁶ is —CH₂OH, —CH₂OAc, —C(═O)OCH₃, —C(═O)CH₃,OCO₂CH₃—OCO₂CH₃, —CH₂NH(CH₃), or —(CH₂)₁₋₂N(CH₃)₂.
 28. The compound ofclaim 1, wherein number of R⁶ groups substituted on the R⁴R⁵ ring isfrom 1 to about
 4. 29. The compound of claim 1, wherein R^(a) and R^(b)are hydrogen, (C₁-C₄)alkyl, aryl or aryl(C₁-C₈)alkylene.
 30. Thecompound of claim 29, wherein R^(a) and R^(b) are hydrogen, methyl orethyl, phenyl or benzyl.
 31. The compound of claim 30, wherein R^(a) is(C₁-C₈)alkyl.
 32. The compound of claim 31, wherein R^(a) is methyl,ethyl, propyl or butyl.
 33. The compound of claim 32, wherein R^(a) is,methyl, ethyl, i-propyl, i-butyl or tert-butyl.
 34. The compound ofclaim 33, wherein R^(a) and R^(b) is a ring.
 35. The compound of claim34, wherein R⁷ is hydrogen, alkyl, aryl or aryl(C₁-C₈)alkylene.
 36. Thecompound of claim 35, wherein R⁷ is hydrogen, methyl or ethyl, phenyl orbenzyl.
 37. The compound of claim 36, wherein R⁷ is H, or methyl. 38.The compound of claim 37, wherein N(R⁷)₂ is amino, methylamino,dimethylamino; ethylamino; pentylamino, diphenylethylamino,pyridylmethylamino, diethylamino or benzylamino.
 39. The compound ofclaim 38, wherein —N(R⁷)₂ is amino, methylamino, dimethylamino;ethylamino; diethylamino or benzylamino.
 40. The compound of claim 39,wherein N(R⁷)₂ is amino, or methylamino.
 41. The compound of claim 40,wherein X is —CH₂OR^(a), —CO₂R^(a), —OC(O)R^(a), —CH₂OC(O)R^(a),—C(O)NR^(a)R^(b).
 42. The compound of claim 41, wherein X is —CH₂OR^(a)or —C(O)NR^(a)R^(b).
 43. The compound of claim 42, wherein X is —CH₂OHor —C(O)NHCH₂CH₃.
 44. The compound of claim 43, wherein m is 0, 1, or 2.45. The compound of claim 44, wherein m is 0, or
 1. 46. The compound ofclaim 18, wherein the rings comprising R⁴, R⁵ and the atom to which theyare connected are selected from the group consisting of:


47. The compound of claim 46, wherein the rings comprising R⁴, R⁵ andthe atom to which they are connected are selected from the groupconsisting of:


48. The compound of claim 47, wherein the ring comprising —C(R³)R⁴R⁵ is2-methylcyclohexane, 2,2-dimethylcyclohexane, 2-phenylcyclohexane,2-ethylcyclohexane, 2,2-diethylcyclohexane, 2-tert-butylcyclohexane,3-methylcyclohexane, 3,3-dimethylcyclohexane, 4-methylcyclohexane,4-ethylcyclohexane, 4-phenyl cyclohexane, 4-tert-butylcyclohexane,4-carboxymethyl cyclohexane, 4-carboxyethyl cyclohexane,3,3,5,5-tetramethyl cyclohexane, 2,4-dimethyl cyclopentane.4-cyclohexanecarboxyic acid, 4-cyclohexanecarboxyic acid esters, or4-methyloxyalkanoyl-cyclohexane.
 49. The compound of claim 48, whereinthe ring comprises —C(R³)R⁴R⁵ is 4-piperidine, 4-piperidene-1-carboxylicacid, 4-piperidine-1-carboxylic acid methyl ester,4-piperidine-1-carboxylic acid ethyl ester, 4-piperidine-1-carboxylicacid propyl ester, 4-piperidine-1-carboxylic acid tert-butyl ester,1-piperidine, 1-piperidine-4-carboxylic acid methyl ester,1-piperidine-4-carboxylic acid ethyl ester, 1-piperidine-4-carboxylicacid propyl ester, 1-piperidine-4-caboxylic acid tert-butyl ester,1-piperidine-4-carboxylic acid methyl ester, 3-piperidine,3-piperidene-1-carboxylic acid, 3-piperidine-1-carboxylic acid methylester, 3-piperidine-1-carboxylic acid tert-butyl ester, 1,4-piperazine,4-piperazine-1-carboxylic acid, 4-piperazine-1-carboxylic acid methylester, 4-piperazine-1-carboxylic acid ethyl ester,4-piperazine-1-carboxylic acid propyl ester, 4-piperazine-1-carboxylicacid tert-butylester, 1,3-piperazine, 3-piperazine-1-carboxylic acid,3-piperazine-1-carboxylic acid methyl ester, 3-piperazine-1-carboxylicacid ethyl ester, 3-piperazine-1-carboxylic acid propyl ester,3-piperidine-1-carboxylic acid tert-butylester,1-piperidine-3-carboxylic acid methyl ester, 1-piperidine-3-carboxylicacid ethyl ester, 1-piperidine-3-carboxylic acid propyl ester or1-piperidine-3-caboxylic acid tert-butyl ester.
 50. The compound ofclaim 49, wherein the ring comprising R⁴ and R⁵ is 2-methyl cyclohexane,2,2-dimethylcyclohexane, 2-phenyl cyclohexane, 2-ethylcyclohexane,2,2-diethylcyclohexane, 2-tert-butyl cyclohexane, 3-methyl cyclohexane,3,3-dimethylcyclohexane, 4-methyl cyclohexane, 4-ethylcyclohexane,4-phenyl cyclohexane, 4-tert-butyl cyclohexane, 4-carboxymethylcyclohexane, 4-carboxyethyl cyclohexane, 3,3,5,5-tetramethylcyclohexane, 2,4-dimethyl cyclopentane, 4-piperidine-1-carboxylic acidmethyl ester, 4-piperidine-1-carboxylic acid tert-butyl ester4-piperidine, 4-piperazine-1-carboxylic acid methyl ester,4-piperidine-1-carboxylic acid tert-butylester,1-piperidine-4-carboxylic acid methyl ester, 1-piperidine-4-caboxylicacid tert-butyl ester, tert-butylester, 1-piperidine-4-carboxylic acidmethyl ester, or 1-piperidine-4-caboxylic acid tert-butyl ester,3-piperidine-1-carboxylic acid methyl ester, 3-piperidine-1-carboxylicacid tert-butyl ester, 3-piperidine, 3-piperazine-1-carboxylic acidmethyl ester, 3-piperidine-1-carboxylic acid tert-butylester,1-piperidine-3-carboxylic acid methyl ester, 1-piperidine-3-caboxylicacid tert-butyl ester.
 51. A compound of claim 1, having the formula:


52. A compound of claim 1, having the formula:


53. A compound of claim 1, having the formula:


54. A compound of claim 1, having the formula:


55. A compound of claim 1, having the formula:


56. A compound of claim 1, having the formula:


57. A compound of claim 1, having the formula:


58. A compound of claim 1, having the formula:


59. A compound of claim 1, having the formula:


60. A compound of claim 1, having the formula:


61. A compound of claim 1, having the formula:


62. A therapeutic method to inhibit an inflammatory response comprisingadministering to a mammal in need of said therapy, an effectiveanti-inflammatory amount of a compound of claim
 1. 63. A therapeuticcomposition comprising a compound of claim 1, in combination with apharmaceutically acceptable carrier.
 64. The composition of claim 63further comprising a Type IV phosphodiesterase inhibitor.
 65. Thecomposition of claim 64 wherein the inhibitor is rolipram.
 66. Thecomposition of claim 65, wherein the carrier is a liquid carrier. 67.The composition of claim 66, which is adapted for parenteral, aerosol ortransdermal administration.
 68. A method for preventing or treating apathological condition or symptom in a mammal, wherein the activity ofA_(A2) adenosine receptors is implicated and agonism of such activity isdesired, comprising administering to said mammal an effective amount ofa compound of claim
 1. 69. The method of claim 68, wherein the mammal isa human.